Haynes Lab:Notebook/CRISPR Editing/2015/03/03

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03/03/2015

Checked concentrations on minipreps from last year of Lonza pMax GFP from Krishna. very low so JW transformed the original along with pMax GFP 6.2 and KAH60.

Sample Read# 260 280 260/280 ng/µL
pMax GFP0.0260.0151.73626.294
pMax GFP0.0230.0131.76623.175
pMax GFP0.0310.0191.63930.851



Ran PCR products from P151/158 on gel. Only single product for each band.

Sequencing:
Need to purify. Will purify 25ul and elute into 35 ul.
Purification results:

Cell type treatment rep 260 280 260/280 ng/µL
Luc14Untreated10.0350.0172.00635.007
Luc14g04610.0320.0162.0231.691
Luc14Untreated20.0450.0222.03444.797
Luc14Untreated30.0460.0241.93846.277
Gal4-EED +doxUntreated10.040.0192.09940.212
Gal4-EED +doxUntreated20.0430.0231.88542.773
Gal4-EED +doxUntreated30.0440.0231.9343.675
Luc14g04620.030.0152.08630.259
Luc14g04630.0310.0152.11331.368
Gal4-EED +doxg04610.0350.0172.04235.122
Gal4-EED +doxg04620.0390.0192.0538.501
Gal4-EED +doxg04630.0410.0221.89741.336
Luc14Untreated10.0320.0161.94931.937
Luc14g04610.030.0151.9429.645


Sent for sequencing at biodesign with P176

Label Cell type treatment rep PCR primer
1Luc14Untreated1P176
2Luc14Untreated2P176
3Luc14Untreated3P176
4Gal4-EED+doxUntreated1P176
5Gal4-EED+doxUntreated2P176
6Gal4-EED+doxUntreated3P176
7Luc14g0341P176
8Luc14g0342P176
9Luc14g0343P176
10Gal4-EED+doxg0341P176
11Gal4-EED+doxg0342P176
12Gal4-EED+doxg0343P176
13Luc14g0461P176
14Luc14g0462P176
15Luc14g0463P176
16Gal4-EED+doxg0461P176
17Gal4-EED+doxg0462P176
18Gal4-EED+doxg0463P176



Ran PCR with P151/158 on g023, g045, g048 PCR with P197/198 diluted 1:1000 from yesterday. Ran on gel.


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