BUGSS:Build-a-BUG:2-3: Difference between revisions
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=== Polymerase chain reaction (PCR) protocol === | === Polymerase chain reaction (PCR) protocol === | ||
* | * Assemble PCR mixture | ||
** PCR master mix - 45.0 μL | ** PCR master mix - 45.0 μL | ||
*** Nuclease-free water -35.6 μL | *** Nuclease-free water -35.6 μL | ||
| Line 23: | Line 23: | ||
*** ''Taq'' DNA polymerase - 0.5 μL | *** ''Taq'' DNA polymerase - 0.5 μL | ||
** Template DNA (3002) - 5.0 μL | ** Template DNA (3002) - 5.0 μL | ||
* | * Amplify in thermal cycler (30 cycles) | ||
** Melt at 95°C for ? sec | ** Melt at 95°C for ? sec | ||
** Anneal at ?°C for ? sec | ** Anneal at ?°C for ? sec | ||
| Line 31: | Line 31: | ||
* [https://www.neb.com/products/m0273-taq-dna-polymerase-with-standard-taq-buffer ''Taq'' DNA polymerase] | * [https://www.neb.com/products/m0273-taq-dna-polymerase-with-standard-taq-buffer ''Taq'' DNA polymerase] | ||
* [http://www.promega.com/products/pcr/routine-pcr/dntp-mix/ Deoxynucleotide triphosphate (dNTP) mix] | * [http://www.promega.com/products/pcr/routine-pcr/dntp-mix/ Deoxynucleotide triphosphate (dNTP) mix] | ||
== Notes == | |||
* | |||
=== Whiteboard === | |||
[[Image:BAB2-2_Agenda.jpg|thumb|left|760px|alt=Build-a-BUG:2-2 agenda|Build-a-BUG:2-2 agenda]] | |||
[[Image:BAB2-2_BioBricks.jpg|thumb|left|760px|alt=BioBrick parts with sizes and restriction enzymes|BioBrick parts with sizes and restriction enzymes]] | |||
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Revision as of 13:28, 18 March 2013
Home Contact Internal Lab Members Projects Protocols Build-a-BUG Build-a-Gene 2016 Build-a-Gene 2015 Build-a-Gene 2014 Build-a-Gene 2013 Chalk Talks
Build-a-BUG: Series 2, Session 3
Saturday, March 9, 2013
Noon to 4:00 PM
This is the third Build-A-BUG workshop in a series of five on yeast. You will learn more about molecular cloning and continue to hone your basic lab techniques, while continuing either your own or somebody else's mating type detector project.
Background reading
Polymerase chain reaction (PCR) protocol
- Assemble PCR mixture
- PCR master mix - 45.0 μL
- Nuclease-free water -35.6 μL
- 10X PCR buffer - 5.0 μL
- Forward primer (VF2) - 1.6 μL
- Reverse primer (VR) - 1.3 μL
- Deoxynucleotide Triphosphate (dNTP) Mix - 1.0 μL
- Taq DNA polymerase - 0.5 μL
- Template DNA (3002) - 5.0 μL
- PCR master mix - 45.0 μL
- Amplify in thermal cycler (30 cycles)
- Melt at 95°C for ? sec
- Anneal at ?°C for ? sec
- Extend at 72°C for ? sec
Materials
Notes
Whiteboard
