Team:Stony Brook/Notebook/Week7
Week 7
7/08:
• Made liquid cultures of all of the transformed SUMO and GST ligated plasmids, MBP vector
• The retransformation of untagged hybrids did not work
7/09:
• Miniprepped the liquid cultures
• Nanodrop of minipreps
• Needed more MBP vector since we had not ligated using MBP vector yet.
7/10:
• Results of MBP vector miniprep
•Made LB and Kanamycin Plates (since our vectors have Kan resistance)
•RE Digest of LnqZA and LnqZE tagged constructs, MBP R1, MBP R2
• Ran RE Digests of MBP on a 1.2% agarose gel
• Ran RE Digests of LnqZA tagged and untagged, lnqZE tagged
• Gel Extract of MBP vectors and inserts that worked
• Made more liquid cultures of all vectors, hybrid inserts
7/11:
• Gel Extraction and nanodrop of inserts
• Miniprep of vectors and hybrid inserts
• RE Digest of Vectors/Inserts
• Ran on gel. All samples split into two gel wells (25uL per well). LnqZA T R1.2 (dropped), MBP 2.1, MBP 3.1, MBP 3.2, LnqZE T and LnqZA U samples did not work.
• Made Liquid cultures of LnqZA U and LnqZE T
7/12:
• Gel Extracted all DNA in gels from RE Digests (7/11)
• Nanodrops Results of Gel Extractions
• Miniprepped and nanodropped LnqZA U and LnqZE T
• Learn about how to prep samples from 7/09 to be sent for sequencing
7/13:
• Prep samples from 7/09 for sequencing!
• Each sample needed to be 8 uL in a 20 uL PCR tube to be accepted
• Each sample must contain 250 ng of DNA
• More vector liquid cultures of GST, MBP, and SUMO
7/14:
• Miniprepped and nanodropped GST, MBP, and SUMO
