Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/27

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Plasmid Verification of DBN001_pSB1A3 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Overview

I'll be performing a restriction digest on the DNA harvested from the DBN001_pSB1A3 ligation & transformation, then running the digestion products on a gel to check size fragments. Hopefully I'll see size fragments that correspond to a successful ligation.

Restriction Digest

Reagent Volume
DNA 600ng (2.5 - 4.7 µL)
10x FastDigest Buffer 3
EcoRI 1
SpeI 1
Water 22.5 - 20.3
Total 30

Digest for 10 minutes at 37°C.

Gel Electrophoresis

Expected size fragments: 2132bp (backbone), 224bp (insert)




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