09/23/2014
Running 2ul of each PCR rxn on a gel, with 3ul of water and 1ul of 6x loading dye.
figuring out volumes of wells so I can know exactly what volume to use for each annealing reaction.
purifying and concentrating all successful PCR rxns using column. should also figure out if 50x loading dye works.
| Sample Read#
|
260
|
280
|
260/280
|
ng/µL
|
ul for 400ng
|
| Luc14 gRNA 20 |
0.141 |
0.075 |
1.864 |
140.539 |
2.85
|
| Luc14 gRNA 21 |
0.176 |
0.098 |
1.788 |
175.643 |
2.28
|
| Luc14 gRNA 22 |
0.198 |
0.106 |
1.869 |
197.974 |
2.02
|
| Luc14 gRNA 27 |
0.131 |
0.07 |
1.868 |
131.014 |
3.05
|
| Luc14 gRNA 29 |
0.159 |
0.086 |
1.858 |
159.485 |
2.51
|
Set up annealing reaction, 10ul total
| '
|
ul for 400ng
|
ul elution solution
|
ul water
|
ul 10x annealing buffer
|
| gRNA 20 |
2.85 |
0.15 |
6 |
1
|
| gRNA 21 |
2.28 |
0.72 |
6 |
1
|
| gRNA 22 |
2.02 |
0.98 |
6 |
1
|
| gRNA 27 |
3.05 |
-0.05 |
6 |
1
|
| gRNA 29 |
2.51 |
0.49 |
6 |
1
|
Added 1ul of enhancer and 1ul of nuclease to each reaction, incubate at 42 for 1 hour.
Ran gel 2ul of ladder, 1ul of each purified PCR reaction (lost a little of gRNA 22), and gRNA 27 unpurified reaction 2 that looked like it didn't work at all.
uncut controls of each gRNA purified pcr of the same volumes as annealing reactions.
| 
Project name
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