User:Matt Hartings/Notebook/Photosynthesis/2012/07/11: Difference between revisions
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==Objective== | ==Objective== | ||
Start cloning sperm whale myoglobin gene into pQE-80 vector | |||
==PCR Cloning== | |||
#Prep primers | |||
##Forward Primer (5'-AACAGGATCCGTTCTGTCTGAAGGTGAATGGCAG-3') 0.3mg MW=10546.9g/mol | |||
###Add 284uL of autoclaved water to forward primer to make 100uM solution | |||
## Reverse Primer (5'-AAGCTTACCCTGGTAACCCAGTTCTTTGTA-3') 0.32mg MW=9132g/mol | |||
###Add 350uL of autoclaved water to reverse primer to make 100uM solution | |||
#Prep PCR tube for reaction | |||
##37uL of autoclaved water | |||
##10uL of 10X Pfusion HF buffer | |||
##1uL of dNTPs | |||
##1uL forward primer (meant to use 0.5uL) | |||
##1uL reverse primer (meant to use 0.5uL) | |||
##1uL of WT swMb DNA | |||
##0.5uL of Pfusion HF enzyme | |||
##50uL of wax | |||
#Place in PCR machine | |||
#Run DEL program. (Tamra had edited an older program) | |||
Revision as of 08:28, 11 July 2012
 Biomaterials Design Lab
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ObjectiveStart cloning sperm whale myoglobin gene into pQE-80 vector PCR Cloning
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