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
|
1 |
P165 |
P198 |
1439
|
2 |
P167 |
P198 |
1406
|
3 |
P151 |
P198 |
1411
|
4 |
P163 |
P198 |
1370
|
5 |
P153 |
P198 |
1329
|
6 |
P149 |
P198 |
1283
|
7 |
P159 |
P198 |
1132
|
8 |
P173 |
P198 |
1087
|
9 |
P161 |
P198 |
1052
|
10 |
P175 |
P198 |
968
|
11 |
P197 |
P158 |
1516
|
12 |
P197 |
P172 |
1496
|
13 |
P197 |
P156 |
1388
|
14 |
P197 |
P170 |
1380
|
15 |
P197 |
P176 |
811
|
16 |
P197 |
P162 |
756
|
17 |
P197 |
P174 |
714
|
18 |
P197 |
P160 |
624
|
19 |
P197 |
P150 |
509
|
20 |
P197 |
P198 |
1660
|
Results all had strong single bands except P156. P156 has a band at the correct size but it's faint.
|