Team:LASATX/Protocols




Protocols

Here is a list of the protocols we used in our project.

STEP 1: PCR Amplification

STEP 2: Gel Electrophoresis

STEP 3: Gel Purification (Promega)

STEP 4: PCR Clean-Up (Zymo) of Remaining PCR Product

STEP 5: Gibson

STEP 6A: Chemical Transformation: One Shot TOP10 Competent Cells

STEP 6B: Electrotransformation of E. coli

STEP 7: Electroporation

STEP 8: ON Cultures

STEP 9: Making Glycerol Stocks

STEP 10: Miniprep (Qiagen)

STEP 11: Sequencing Sample Submission

STEP 12: Analyze Sequences, Sequence Verification

STEP 13: Quikchange

STEP 14: PCR Transformation w/ Dpn1

Restriction Digest

Ligation

Sequence Verification: TOPO-TA

pchBA Genomic Extraction

Assembly of Entire COWg Construct

Experimentation with Full Construct