BME100 s2015:Group13 12pmL4: Difference between revisions
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| [[Image: | | [[Image:montina.jpg|100px|thumb|Name: Tina Monteilh<br>Role(s)]] | ||
| [[Image:Trisha | | [[Image:Trisha.jpg|100px|thumb|Name:Trisha Dasgupta<br>Role(s)]] | ||
| [[Image:Riley | | [[Image:Riley.jpg|100px|thumb|Name: student<br>Role(s)]] | ||
| [[Image:Payson | | [[Image:Payson.jpg|100px|thumb|Name:Payson Wallach<br>Role(s)]] | ||
| [[Image:Robel | | [[Image:Robel Okbe.jpg|100px|thumb|Name: Robel Okbe<br>Role(s)]] | ||
=LAB 4 WRITE-UP= | =LAB 4 WRITE-UP= | ||
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'''DNA Sample Set-up Procedure''' | '''DNA Sample Set-up Procedure''' | ||
<!-- In your own words, write an easy-to-comprehend list of the steps your group took to set up the PCR reaction in the PCR reaction tubes. End with placing the tubes in to the thermal cycler. Do not copy-paste the instructions from the Workbook. That will be considered plagiarism. --> | <!-- In your own words, write an easy-to-comprehend list of the steps your group took to set up the PCR reaction in the PCR reaction tubes. End with placing the tubes in to the thermal cycler. Do not copy-paste the instructions from the Workbook. That will be considered plagiarism. --> | ||
# | # Collect all needed Materials | ||
# | # Label the sides of the PCR tubes with the Tube labels listed above. | ||
# | # Place tubes in rack | ||
# Micropipette 50 microliters of the positive control mix into the empty tube labelled positive control | |||
# Repeat step 5 for the negative control, patient 1 replicates 1,2, 3, and for patient 2 replicates, 1, 2, 3 | |||
# Make sure the lids are tightly closed | |||
# Place tubes into the assigned PCR machine, make sure to appropriately label the side of the PCR machine your group's tubes were placed on for easy identification later on | |||
'''OpenPCR program''' | '''OpenPCR program''' | ||
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Template DNA is used to make the complementary strand. Primers are used to help copy the DNA by attaching to sites on the DNA strands that are at either end of the segment that needs to be copied. Taq polymeraseis an enzyme that assembles new DNA from nucleotides. Deoxyribonucleotides (dNTPs) are the building blocks from which the DNA polymerase (Taq polymerase) uses to make the new DNA strand. <br> | Template DNA is used to make the complementary strand. Primers are used to help copy the DNA by attaching to sites on the DNA strands that are at either end of the segment that needs to be copied. Taq polymeraseis an enzyme that assembles new DNA from nucleotides. Deoxyribonucleotides (dNTPs) are the building blocks from which the DNA polymerase (Taq polymerase) uses to make the new DNA strand. <br> | ||
Q2<br> | Q2<br> | ||
During the initial step, the system is heated up to 95 degrees Celsius which allows for all of the components to heat up. During the denature phase, which is also at 95 degrees Celsius for 30 seconds the template DNA experiences DNA melting which makes single-stranded DNA. During the anneal phase which is at 57 degrees Celsius for 30 seconds the template the primers are able to attach to the single stranded DNA template. The extension phase takes place at 72 degrees Celsius for 30 seconds, and the Taq polymerase is at its optimum activity level allowing it to assemble a new strand of DNA along with a complementary strand using the deoxyribonucleotides. The final step takes place at 72 degrees Celsius for 3 minutes, and this is just to ensure that all of the single-stranded DNA was extended. The final hold occurs at 4 degrees Celsius and this is for short-term storage of the reaction. | |||
<br> | |||
Q3 <br> | Q3 <br> | ||
Adenine pairs with Thymine while Cytosine pairs with Guanine. <br> | Adenine pairs with Thymine while Cytosine pairs with Guanine. <br> | ||
Q4<br> | Q4<br> | ||
Base-pairing occurs during the extension at 72 degrees Celsius for 30 seconds since at this point a new strand of DNA is being assembled by attaching to the complementary strand. This only occurs through the base pairing. Base pairing also occurs at the anneal phase since the primers must attach to the DNA template, and this probably occurs due to base pairing. | |||
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What is a nucleotide? The organic base units of DNA and RNA | What is a nucleotide? The organic base units of DNA and RNA | ||
What is a polymorphism? A point mutation that is expressed in multiple forms within a population. | What is a polymorphism? A point mutation that is expressed in multiple forms within a population. | ||
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Disease reverse primer (20 nt): gaaacaccagggctcagggt | Disease reverse primer (20 nt): gaaacaccagggctcagggt | ||
'''Primer Design and Testing''' | '''Primer Design and Testing''' | ||
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<br><br> | <br><br> |
Latest revision as of 23:52, 31 March 2015
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