BUGSS:Build-a-BUG:2-3
From OpenWetWare
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