01/09/2015
| Sample Read#
|
260
|
280
|
260/280
|
ng/µL
|
| 31 |
0.114 |
0.061 |
1.854 |
113.641
|
| 35 |
0.097 |
0.051 |
1.913 |
97.073
|
| 38 |
0.117 |
0.062 |
1.887 |
117.472
|
| 40 |
0.103 |
0.055 |
1.89 |
103.387
|
| 41 |
0.128 |
0.067 |
1.907 |
128.36
|
| 42 |
0.102 |
0.055 |
1.855 |
101.652
|
| 48 |
0.109 |
0.06 |
1.806 |
108.503
|
| 49 |
0.096 |
0.051 |
1.871 |
95.959
|
| Sample Read#
|
ng/µL
|
ul for 400ng
|
ulH2O
|
ul ann buff
|
| 31 |
113.641 |
3.52 |
9.98 |
1.5
|
| 35 |
97.073 |
4.12 |
9.38 |
1.5
|
| 38 |
117.472 |
3.41 |
10.09 |
1.5
|
| 40 |
103.387 |
3.87 |
9.63 |
1.5
|
| 41 |
128.36 |
3.12 |
10.38 |
1.5
|
| 42 |
101.652 |
3.93 |
9.57 |
1.5
|
| 48 |
108.503 |
3.69 |
9.81 |
1.5
|
| 49 |
95.959 |
4.17 |
9.33 |
1.5
|
Had 2-15ul annealing reactions for all 22 gRNA treated Luc14 samples. Added 1ul Surveyor enzyme and 1ul of enhancer (mixed together first) to each +nuc tube and 2ul of elution solution to the -nuc tubes. Incubated at 42degC for 45 minutes, added 1.7ul stop solution to every tube.
Analysis with bioanalyzer
Made 1:20 dilutions of each Surveyor reaction. Ran one ul on each chip. Ran untreated samples +/-Luc on every chip. Data looks really solid, can select some gRNAs to proceed with.
Analysis with 2% agarose gel
Added 3.74ul 6x loading dye to the remaining 17.7ul of surveyor reaction. Ran on a large 2% agarose gel. Made the gel very thick so I could fit all the reaction in the well but some still spilled out. I also took a long time to load the gel so I think that messed up some of the early lanes. It looks like there are two bands in those lanes, even in the ladder. I think maybe I ran it too fast (100V). I need to learn more about 2% gels. The bands are consistent with the bioanalyzer data but it's not close to publishable data.
Thawed Luc14 gal4-eed cells for experiments
| 
Project name
Main project page
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