Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/15

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Transformation Results for DBN001_pSB1A3 Main project page
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Overview

Here are the colonies observed after one weekend grown at room temperature. I'll also be doing a plasmid miniprep of mCh_pSB1A3 so that I have more stocks of the cloning vector.

Plate # colonies
positive controlmany dozens
sample2
negative control2

I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.

mCh_pSB1A3 Plasmid Miniprep

Decent yield for all four colonies picked.

--DNA Concentration--

Sample ID Concentration (ng/µL) St Dev (ng/µL)
mCh Vector 1392.115.5
mCh Vector 2315.518.1
mCh Vector 3319.76.3
mCh Vector 4375.17.1

--Restriction Digest--

Reagent Volume
DNA600ng (1.9 - 1.5 µL)
10x FastDigest Buffer3
EcoRI1
PstI1
Water23
Total30

Digest for 15 minutes at 37°C.



--Gel Electrophoresis--

Image:2015-04-15_mCherry_vector_digest_annotated.png

Expected size fragments from EcoRI/PstI digest: 2114, 980



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