Daniel:Notebook/HiResChrPaint/2013-9-4
Jump to navigation
Jump to search
BSPP Control (Started 09/03/2013)[edit]
DpnII Digestion (Started 09/03/2013)[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 | 127.7 | 130.6 | 110.1 | 109.3 |
ug in 30 uL | 3.8 | 3.9 | 3.3 | 3.3 |
total | 7.7 | 6.6 |
TBE Gel[edit]
1. Lanes; (5.5 uL TBE; 2 uL 6x buffer, 0.5 uL sample; load 7 uL for all)
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 25bp ladder | BSPP 1AB | BSPP 1CD | BSPP 2AB | BSPP 2CD |
2. Run for 30 minutes at 200V 3. Stain with 3 uL SYBR gold for 2 minutes
Gel Image
File:2013-09-04-DpnIIDigest-BSPP.jpg
The gel looks fine, so I'm going to proceed as usual.
Lambda Exonuclease Digestion[edit]
1. Reaction table
Sample | BSPP 1AB | BSPP 1CD | BSPP 2AB | BSPP 2CD |
ng/uL | 127.7 | 130.6 | 110.1 | 109.3 |
ug in 25 uL | 3.2 | 3.3 | 2.8 | 2.7 |
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 5. Measure Concentration in Nanodrop
ssNanodrop Results[edit]
ssProbe | BSPP 1 | BSPP 2 |
ng/uL | 119 | 100.3 |
ug in 20 uL | 2.4 | 2.0 |