Daniel:Notebook/HiResChrPaint/2014-1-9: Difference between revisions
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===TBE Gel=== | ===TBE Gel=== | ||
1. '''For Each Sample''': | 1. '''For Each Sample''': 2 uL dye; load 7.2 uL | ||
{{GelLanes|Lane3=10bp ladder|Lane5=Alu2-B Pre-Digest|Lane7=Alu2-B Post-Digest|Lane9=Alu2-AB Post-Digest}} | |||
{{GelLanes|Lane3=0.5 uL sample, 5.5 TBE|Lane5=1 uL sample, 5 TBE|Lane7=1 uL sample, 5 TBE|Lane9=0.5 uL sample, 5.5 TBE}} | |||
2. Run gel for 27 minutes at 220V | 2. Run gel for 27 minutes at 220V | ||
3. Stain 2 minutes with 3 uL SYBR gold | 3. Stain 2 minutes with 3 uL SYBR gold | ||
'''Gel Image''' | '''Gel Image''' |
Revision as of 21:28, 9 January 2014
Alu Control (Started 1/08/2014)
DpnII Digestion
I'm going to do a test digestion/gel to make sure this works. Since I had such low input I'm going to use 2 samples and pool after the digestion.
1. Reaction Table
Sample | ALU2-A | ALU2-B |
ng/uL | 25.1 | 25 |
ug in 35 uL | 0.9 | 0.9 |
uL 10X buffer | 5 | 5 |
uL DpnII (50U/uL) | 1 | 1 |
uL nfH2O | 9 | 9 |
Total | 50 | 50 |
2. Incubate samples at 37C for 2 hours; 20 minute heat shock at 65C 3. Column Purification, elute with 30 uL EB 4. Measure concentrations in nanodrop
Nanodrop Results
ALU2-AB: 37.3 ng/uL; 1.1 ug total
TBE Gel
1. For Each Sample: 2 uL dye; load 7.2 uL
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 10bp ladder | Alu2-B Pre-Digest | Alu2-B Post-Digest | Alu2-AB Post-Digest |
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 0.5 uL sample, 5.5 TBE | 1 uL sample, 5 TBE | 1 uL sample, 5 TBE | 0.5 uL sample, 5.5 TBE |
2. Run gel for 27 minutes at 220V 3. Stain 2 minutes with 3 uL SYBR gold
Gel Image