Team:Nevada/Week 7

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Week 7: July 2 - July 6

Contents

July 2

Jasmine, Joe, and Michelle:
Amplified CP using PCR (adjusted amount of template)
Digested EP* with SpeI and NsiI
Ligated RFP* and EP*
Jeremiah & Chris:
Colony PCR of SBP-TBP in ExV (colonies 4 & 5 may be positive)
Cultured LRP
Digested TBP++ with SpeI and PstI

July 3

Jasmine, Joe, and Michelle:
Digested CP with EcoRI and SpeI
Digested EP* with EcoRI and SpeI
Amplified more CP using PCR
Transformed RFP*-EP* ligation and plated onto Amp plate
Justin and Dafne
Miniprep SBP-B12(in expression plasmid) colonies
Digest with Spe I to ensure the correct plasmid size
Resulting electrophoresis analysis indicated successful creation of SBP-B12 in expression plasmid construct

July 4

Jeremiah & Chris:
Both standard and Phusion ligation of SBP-TBPa EXVTransform LRP-TBP++


July 5

Jasmine and Joe:
Checked 8 RFP*-EP* colonies with colony PCR
Cultured colonies 7 and 8
Amplified more RFP* using PCR
Digested CP with EcoRI and SpeI (no purification after PCR)
Gel purified EP*digested with EcoRI and SpeI
Jeremiah & Chris
Gel purified EP*digested with EcoRI and SpeI
aDigest SBP-TBP for expression (again)with XbaI and PstI


July 6

Jasmine and Joe:
Dephosphorylated EP*
Ligated CP and EP*
Transformed into competent cells and plated onto Amp plate