Team:Nevada/Week 2

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Week 2: May 28 - June 1

Contents

May 28

Michelle:
Ligation of SBP + LRP transformed onto Kanamyacin (KAN) plates. Gel analysis of SBP.
Justin and Dafne:
Digest SBP and B12 again.
Initial digests were unsuccessful

May 29

Jeremiah and Chris:
Made new CCMB80 buffer for use in purification and new competent cells in SOB. Re-ligated SBP-TBP as gel showed no results for first attempt.
Justin and Dafne:
Ligate B12 into SBP
SBP was dephosphorylated first, using Antarctic phosphatase
Michelle and Joe:
Check transformation of SBP + LRP from KAN plates. Made more competent cells.

May 30

Michelle and Joe:
Test competent cells and prepared for transformation of RFP in the future.
Jeremiah and Chris:
Testing for contamination in digests

May 31

Michelle and Joe:
Miniprep and digest LRP with PstI and XbaI and SBP with SpeI and PstI. Ran gel of LRP and SBP digestions.
Colony PCR followed by ligation of SBP (SpeI and PstI) with LRP (PstI and XbaI).
Transformation of RFP into competent cells.
Jeremiah and Chris:
Colony PCR done of ligated and transformed colonies of SBP-TBP
Justin and Dafne:
Digest plasmids again. Initial digests were unsuccessful

June 1

Michelle and Joe:
Transformation of SBP (SpeI and PstI) and LRP (PstI and XbaI) ligation onto KAN plates.
Jeremiah and Chris:
Competent cells tested to ensure transformability – did not work.
Transformed SBP-TBP into competent cells