Haynes Lab:Notebook/CRISPR Editing/2015/01/14

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01/14/2015

Picked colonies from 6 of the transformation plates from yesterday into 2-6mL cultures to miniprep tomorrow.

Testing PCR primers
I have a ton of primers that amplify across my target sequence. I tested all of them by pairing all the forward primers with P198 and all the reverse primers with P197, both of which I know work very well.

95 for 3min
30 cycles
95 for 30s
60 for 30s
72 for 2 min
72 for 10min
' F primer R primer exp length
1P165P1981439
2P167P1981406
3P151P1981411
4P163P1981370
5P153P1981329
6P149P1981283
7P159P1981132
8P173P1981087
9P161P1981052
10P175P198968
11P197P1581516
12P197P1721496
13P197P1561388
14P197P1701380
15P197P176811
16P197P162756
17P197P174714
18P197P160624
19P197P150509
20P197P1981660

Results
all had strong single bands except P156. P156 has a band at the correct size but it's faint.


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