Daniel:Notebook/HiResChrPaint/2013-10-23
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BSPP Control (Started 10/22/2013)[edit]
DpnII Digestion (Started 10/22/2013)[edit]
3. Column Purification, elute with 30 uL EB 4. Measure concentrations in nanodrop
Nanodrop Results[edit]
Sample | BSPP 1AB | BSPP 1CD | BSPP 2AB | BSPP 2CD |
ng/uL | 166.5 | 168.8 | 180.2 | 143 |
ug in 30 uL | 5.0 | 5.1 | 5.4 | 4.3 |
total | 10.1 | 9.7 |
TBE Gel[edit]
1. Lanes; (5.5 uL TBE; 2 uL 6x buffer, 0.5 uL sample; load 7.2 uL for all)
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 10bp ladder | BSPP 1AB | BSPP 1CD | BSPP 2AB | BSPP 2CD |
2. Run for 23 minutes at 220V 3. Stain with 3 uL SYBR gold for 2 minutes
Gel Image
Lambda Exonuclease Digestion[edit]
1. Reaction table
Sample | BSPP 1AB | BSPP 1CD | BSPP 2AB | BSPP 2CD |
ng/uL | 166.5 | 168.8 | 180.2 | 143 |
ug in 25 uL | 4.2 | 4.2 | 4.5 | 3.6 |
uL added | 25 | 25 | 25 | 25 |
uL 10x buffer | 5 | 5 | 5 | 5 |
uL Lambda Exonuclease (5U/uL) | 3 | 3 | 3 | 3 |
uL nfH2O | 17 | 17 | 17 | 17 |
total | 50 | 50 | 50 | 50 |
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C 3. Pool samples 4. ssDNA column purification, elute with 20 uL nfH2O 5. Measure Concentration in Nanodrop