http://openwetware.org/index.php?title=PCR_techniques&feed=atom&action=historyPCR techniques - Revision history2013-05-18T12:35:50ZRevision history for this page on the wikiMediaWiki 1.13.2http://openwetware.org/index.php?title=PCR_techniques&diff=573512&oldid=prevChristopher C Vanlang at 10:30, 22 December 20112011-12-22T10:30:12Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[PCR inhibitors]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[PCR inhibitors]]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]] [[Category:RNA]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]] [[Category:RNA<ins class="diffchange diffchange-inline">]] [[Category:PCR</ins>]]</div></td></tr>
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</table>Christopher C Vanlanghttp://openwetware.org/index.php?title=PCR_techniques&diff=430313&oldid=prevJakob Suckale: links2010-07-07T11:59:05Z<p>links</p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**qRT-PCT can be done either in [[qRT-PCR/Single tube|one tube]] or [[qRT-PCR/Two tubes|two tubes]].</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**qRT-PCT can be done either in [[qRT-PCR/Single tube|one tube]] or [[qRT-PCR/Two tubes|two tubes]].</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>=== <del class="diffchange diffchange-inline">Links </del>===</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>=== <ins class="diffchange diffchange-inline">See also </ins>===</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [http://en.wikipedia.org/wiki/Polymerase_chain_reaction#Practical_modifications_to_the_PCR_technique PCR on wikipedia]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [http://en.wikipedia.org/wiki/Polymerase_chain_reaction#Practical_modifications_to_the_PCR_technique PCR on wikipedia]</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[PCR]] - how to do a standard PCR</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[PCR]] - how to do a standard PCR</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[Designing primers]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[Designing primers]]</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[Mouse tissue lysis for genotyping]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[Mouse tissue lysis for genotyping]]</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">* [[PCR inhibitors]]</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]] [[Category:RNA]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]] [[Category:RNA]]</div></td></tr>
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</table>Jakob Suckalehttp://openwetware.org/index.php?title=PCR_techniques&diff=430269&oldid=prevJakob Suckale: hub icon2010-07-07T09:42:27Z<p>hub icon</p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">{{back to protocols}}</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[Image:Hub icon.png|right]]</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods with links to individual pages for detailed information.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods with links to individual pages for detailed information.</div></td></tr>
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</table>Jakob Suckalehttp://openwetware.org/index.php?title=PCR_techniques&diff=288256&oldid=prevTorsten Waldminghaus at 17:28, 23 February 20092009-02-23T17:28:20Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods with links to individual pages for detailed information.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods with links to individual pages for detailed information.</div></td></tr>
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</table>Torsten Waldminghaushttp://openwetware.org/index.php?title=PCR_techniques&diff=157212&oldid=prevReshma P. Shetty at 21:22, 9 October 20072007-10-09T21:22:39Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of starting nucleic acid template. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''[[real-time PCR]]'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis (qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish a linear relationship between band intensity and starting material, this approach is also quantitative.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of starting nucleic acid template. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''[[real-time PCR]]'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis (qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish a linear relationship between band intensity and starting material, this approach is also quantitative.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[Real-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[Real-time PCR]] is used to determine the quantity of DNA <ins class="diffchange diffchange-inline">during the PCR step (i.e. in "</ins>real-time<ins class="diffchange diffchange-inline">") </ins>and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[qRT-PCR|Quantitative RT-PCR]] ('''q'''uantitative '''r'''everse '''t'''ranscription PCR) refers to quantitative PCR of cDNA (which has been reverse transcribed from RNA). Like quantitative PCR, you can quantify the starting RNA template either during the reaction ("real-time reverse transcription PCR") or at the end of the PCR step.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[qRT-PCR|Quantitative RT-PCR]] ('''q'''uantitative '''r'''everse '''t'''ranscription PCR) refers to quantitative PCR of cDNA (which has been reverse transcribed from RNA). Like quantitative PCR, you can quantify the starting RNA template either during the reaction ("real-time reverse transcription PCR") or at the end of the PCR step.</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">**qRT-PCT can be done either in [[qRT-PCR/Single tube|one tube]] or [[qRT-PCR/Two tubes|two tubes]].</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td></tr>
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</table>Reshma P. Shettyhttp://openwetware.org/index.php?title=PCR_techniques&diff=157200&oldid=prevReshma P. Shetty at 21:13, 9 October 20072007-10-09T21:13:15Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods with links to individual pages for detailed information.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods with links to individual pages for detailed information.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[PCR]] <del class="diffchange diffchange-inline">or </del>'''p'''olymerase '''c'''hain '''r'''eaction is a method for exponentially amplifying a fragment of DNA in vitro.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[PCR]] <ins class="diffchange diffchange-inline">(</ins>'''p'''olymerase '''c'''hain '''r'''eaction<ins class="diffchange diffchange-inline">) </ins>is a method for exponentially amplifying a fragment of DNA in vitro.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Nested PCR]] is 2 successive PCRs with the 2nd set of primers nested inside the 1st pair. It is used to reduce unspecific products.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Nested PCR]] is 2 successive PCRs with the 2nd set of primers nested inside the 1st pair. It is used to reduce unspecific products.</div></td></tr>
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</table>Reshma P. Shettyhttp://openwetware.org/index.php?title=PCR_techniques&diff=157199&oldid=prevReshma P. Shetty at 21:12, 9 October 20072007-10-09T21:12:57Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods <del class="diffchange diffchange-inline">linking </del>to individual pages for detailed information.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction ([[PCR]]). Below is an overiew of important PCR methods <ins class="diffchange diffchange-inline">with links </ins>to individual pages for detailed information.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[PCR]] or '''p'''olymerase '''c'''hain '''r'''eaction is a method for exponentially amplifying a fragment of DNA in vitro.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[PCR]] or '''p'''olymerase '''c'''hain '''r'''eaction is a method for exponentially amplifying a fragment of DNA in vitro.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Colony PCR]] is a PCR technique to detect DNA in bacterial colonies. Colonies are picked, lysed, and amplified by PCR. It is used to detect successful ligations or recombinations among large numbers of bacterial colonies.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Colony PCR]] is a PCR technique to detect DNA in bacterial colonies. Colonies are picked, lysed, and amplified by PCR. It is used to detect successful ligations or recombinations among large numbers of bacterial colonies.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[RT-PCR]] ('''<del class="diffchange diffchange-inline">R</del>'''everse '''<del class="diffchange diffchange-inline">T</del>'''ranscription PCR) is essentially normal PCR preceded by a reverse transcription converting RNA to cDNA. It is used to amplify an RNA molecule like messenger RNAs. Confusingly, RT-PCR is used as an abbreviation for real-time PCR.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[RT-PCR]] ('''<ins class="diffchange diffchange-inline">r</ins>'''everse '''<ins class="diffchange diffchange-inline">t</ins>'''ranscription PCR) is essentially normal PCR preceded by a reverse transcription converting RNA to cDNA. It is used to amplify an RNA molecule like messenger RNAs. Confusingly, RT-PCR is used as an abbreviation for real-time PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of starting nucleic acid template. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''[[real-time PCR]]'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis (qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish a linear relationship between band intensity and starting material, this approach is also quantitative.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of starting nucleic acid template. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''[[real-time PCR]]'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis (qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish a linear relationship between band intensity and starting material, this approach is also quantitative.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Real-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Real-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[qRT-PCR|Quantitative RT-PCR]] <del class="diffchange diffchange-inline">or </del>'''q'''uantitative '''r'''everse '''t'''ranscription PCR refers to quantitative PCR of cDNA (which has been reverse transcribed from RNA). Like quantitative PCR, you can quantify the starting RNA template either during the reaction ("real-time reverse transcription PCR") or at the end of the PCR step.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[qRT-PCR|Quantitative RT-PCR]] <ins class="diffchange diffchange-inline">(</ins>'''q'''uantitative '''r'''everse '''t'''ranscription PCR<ins class="diffchange diffchange-inline">) </ins>refers to quantitative PCR of cDNA (which has been reverse transcribed from RNA). Like quantitative PCR, you can quantify the starting RNA template either during the reaction ("real-time reverse transcription PCR") or at the end of the PCR step.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td></tr>
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</table>Reshma P. Shettyhttp://openwetware.org/index.php?title=PCR_techniques&diff=157197&oldid=prevReshma P. Shetty at 21:11, 9 October 20072007-10-09T21:11:35Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Real-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Real-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[qRT-PCR|Quantitative RT-PCR]] or '''q'''uantitative '''r'''everse '''t'''ranscription PCR refers to quantitative PCR of cDNA (which has been reverse transcribed from RNA). Like quantitative PCR, you can quantify the starting RNA template either during the reaction ("<del class="diffchange diffchange-inline">Real</del>-time reverse transcription PCR") or at the end of the PCR step.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[qRT-PCR|Quantitative RT-PCR]] or '''q'''uantitative '''r'''everse '''t'''ranscription PCR refers to quantitative PCR of cDNA (which has been reverse transcribed from RNA). Like quantitative PCR, you can quantify the starting RNA template either during the reaction ("<ins class="diffchange diffchange-inline">real</ins>-time reverse transcription PCR") or at the end of the PCR step.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[Mouse tissue lysis for genotyping]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* [[Mouse tissue lysis for genotyping]]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:In vitro]] [[Category:DNA<ins class="diffchange diffchange-inline">]] [[Category:RNA</ins>]]</div></td></tr>
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</table>Reshma P. Shettyhttp://openwetware.org/index.php?title=PCR_techniques&diff=157196&oldid=prevReshma P. Shetty at 21:09, 9 October 20072007-10-09T21:09:54Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction (PCR). Below is an overiew of important PCR methods linking to individual pages for detailed information.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Several techniques have have been derived from the basic polymerase chain reaction (<ins class="diffchange diffchange-inline">[[</ins>PCR<ins class="diffchange diffchange-inline">]]</ins>). Below is an overiew of important PCR methods linking to individual pages for detailed information.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[<del class="diffchange diffchange-inline">nested </del>PCR]] is <del class="diffchange diffchange-inline">2 successive PCRs with the 2nd set </del>of <del class="diffchange diffchange-inline">primers nested inside the 1st pair. It is used to reduce unspecific products</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[PCR]] <ins class="diffchange diffchange-inline">or '''p'''olymerase '''c'''hain '''r'''eaction </ins>is <ins class="diffchange diffchange-inline">a method for exponentially amplifying a fragment </ins>of <ins class="diffchange diffchange-inline">DNA in vitro</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[<del class="diffchange diffchange-inline">multiplex </del>PCR]] is <del class="diffchange diffchange-inline">a PCR </del>with <del class="diffchange diffchange-inline">>1 primer </del>pair <del class="diffchange diffchange-inline">run in a single reaction</del>. It <del class="diffchange diffchange-inline">reduced material consumption but </del>is <del class="diffchange diffchange-inline">hard </del>to <del class="diffchange diffchange-inline">optimise. It is often used in mouse genotyping for example</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[<ins class="diffchange diffchange-inline">Nested </ins>PCR]] is <ins class="diffchange diffchange-inline">2 successive PCRs </ins>with <ins class="diffchange diffchange-inline">the 2nd set of primers nested inside the 1st </ins>pair. It is <ins class="diffchange diffchange-inline">used </ins>to <ins class="diffchange diffchange-inline">reduce unspecific products</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[<del class="diffchange diffchange-inline">colony </del>PCR]] is a PCR <del class="diffchange diffchange-inline">technique to detect DNA </del>in <del class="diffchange diffchange-inline">bacterial colonies</del>. <del class="diffchange diffchange-inline">Colonies are picked, lysed, and amplified by PCR</del>. It is used <del class="diffchange diffchange-inline">to detect successful ligations or recombinations among large numbers of bacterial colonies</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[<ins class="diffchange diffchange-inline">Multiplex </ins>PCR]] is a PCR <ins class="diffchange diffchange-inline">with >1 primer pair run </ins>in <ins class="diffchange diffchange-inline">a single reaction</ins>. <ins class="diffchange diffchange-inline">It reduced material consumption but is hard to optimise</ins>. It is <ins class="diffchange diffchange-inline">often </ins>used <ins class="diffchange diffchange-inline">in mouse genotyping for example</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[RT-PCR]] ('''R'''everse '''T'''ranscription PCR) is essentially normal PCR preceded by a reverse transcription converting RNA to cDNA. It is used to <del class="diffchange diffchange-inline">indirectly detect/measure RNAs, e.g. </del>messenger RNAs. Confusingly, RT PCR is used as an abbreviation for real-time PCR.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*[[Colony PCR]] is a PCR technique to detect DNA in bacterial colonies. Colonies are picked, lysed, and amplified by PCR. It is used to detect successful ligations or recombinations among large numbers of bacterial colonies.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[RT-PCR]] ('''R'''everse '''T'''ranscription PCR) is essentially normal PCR preceded by a reverse transcription converting RNA to cDNA. It is used to <ins class="diffchange diffchange-inline">amplify an RNA molecule like </ins>messenger RNAs. Confusingly, RT<ins class="diffchange diffchange-inline">-</ins>PCR is used as an abbreviation for real-time PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of starting nucleic acid template. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''[[real-time PCR]]'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis (qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish a linear relationship between band intensity and starting material, this approach is also quantitative.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of starting nucleic acid template. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''[[real-time PCR]]'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis (qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish a linear relationship between band intensity and starting material, this approach is also quantitative.</div></td></tr>
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</table>Reshma P. Shettyhttp://openwetware.org/index.php?title=PCR_techniques&diff=157195&oldid=prevReshma P. Shetty at 21:06, 9 October 20072007-10-09T21:06:00Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[RT-PCR]] ('''R'''everse '''T'''ranscription PCR) is essentially normal PCR preceded by a reverse transcription converting RNA to cDNA. It is used to indirectly detect/measure RNAs, e.g. messenger RNAs. Confusingly, RT PCR is used as an abbreviation for real-time PCR.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*[[RT-PCR]] ('''R'''everse '''T'''ranscription PCR) is essentially normal PCR preceded by a reverse transcription converting RNA to cDNA. It is used to indirectly detect/measure RNAs, e.g. messenger RNAs. Confusingly, RT PCR is used as an abbreviation for real-time PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of <del class="diffchange diffchange-inline">DNA</del>. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''real-time <del class="diffchange diffchange-inline">PCRs</del>'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis<del class="diffchange diffchange-inline">, but since this is just </del>a <del class="diffchange diffchange-inline">rough but quick estimate</del>, <del class="diffchange diffchange-inline">many researches don't consider </del>this <del class="diffchange diffchange-inline">a Q-PCR</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[Q-PCR]] ('''q'''uantitative PCR) is used to determine the quantity of <ins class="diffchange diffchange-inline">starting nucleic acid template</ins>. There are 2 main methods: PCR with dsDNA dyes and PCR with probes. dsDNA dyes and probes allow measurement while the PCR is running, i.e. in real-time, and are therefore often named ''<ins class="diffchange diffchange-inline">[[</ins>real-time <ins class="diffchange diffchange-inline">PCR]]</ins>'' (see below). Probes are superior to dsDNA dyes but also more expensive. DNA can also be quantified with a simple PCR followed by agarose gel electrophoresis <ins class="diffchange diffchange-inline">(qPCR end-point assay). If done properly with serial dilutions of starting DNA template to establish </ins>a <ins class="diffchange diffchange-inline">linear relationship between band intensity and starting material</ins>, this <ins class="diffchange diffchange-inline">approach is also quantitative</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">:</del>*[[<del class="diffchange diffchange-inline">real</del>-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR<del class="diffchange diffchange-inline">, </del>which overlaps with reverse transcription PCR<del class="diffchange diffchange-inline">; QRT-PCR or RTQ-PCR are better acronyms</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[<ins class="diffchange diffchange-inline">Real</ins>-time PCR]] is used to determine the quantity of DNA real-time and thus a subset of the Q-PCR methods. It uses fluorescent dyes or fluorophore-DNA probes to measure the amount of amplification in real time. This is used to infer starting amount. Real-time PCR is often confusingly abbreviated as RT PCR which overlaps with reverse transcription PCR.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">::</del>*[[qRT-PCR|<del class="diffchange diffchange-inline">quantitative </del>RT-PCR]] or ''<del class="diffchange diffchange-inline">real-time</del>'' <del class="diffchange diffchange-inline">RT-</del>PCR <del class="diffchange diffchange-inline">is a special type of real-time </del>PCR <del class="diffchange diffchange-inline">with </del>reverse transcribed RNA <del class="diffchange diffchange-inline">as template</del>. <del class="diffchange diffchange-inline">Basically</del>, <del class="diffchange diffchange-inline">RT + Q</del>-PCR. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*[[qRT-PCR|<ins class="diffchange diffchange-inline">Quantitative </ins>RT-PCR]] or '''<ins class="diffchange diffchange-inline">q</ins>'<ins class="diffchange diffchange-inline">''uantitative '''r'''everse '''t'''ranscription </ins>PCR <ins class="diffchange diffchange-inline">refers to quantitative </ins>PCR <ins class="diffchange diffchange-inline">of cDNA (which has been </ins>reverse transcribed <ins class="diffchange diffchange-inline">from </ins>RNA<ins class="diffchange diffchange-inline">)</ins>. <ins class="diffchange diffchange-inline"> Like quantitative PCR</ins>, <ins class="diffchange diffchange-inline">you can quantify the starting RNA template either during the reaction ("Real</ins>-<ins class="diffchange diffchange-inline">time reverse transcription PCR") or at the end of the </ins>PCR <ins class="diffchange diffchange-inline">step</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Links ===</div></td></tr>
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</table>Reshma P. Shetty