Daniel:Notebook/HiResChrPaint/2014-1-9: Difference between revisions

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>Djacobse
>Djacobse
Line 57: Line 57:
===TBE Gel===
===TBE Gel===


  1. '''For Each Sample''': 1 uL sample, 5 uL TBE, 2 uL dye; load 7.2 uL
  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|Lane5=Alu2-AB Post-Digest}}


{{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)

Back to Calendar

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