Daniel:Notebook/HiResChrPaint/2013-10-23: Difference between revisions
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1. Reaction table | 1. Reaction table | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" align="center" | |||
| width="65" height="15" | Sample | |||
| width="65" | BSPP 1AB | |||
| width="65" | BSPP 1CD | |||
| width="65" | BSPP 2AB | |||
| width="65" | BSPP 2CD | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ng/uL | |||
| align="center" | 166.5 | |||
| align="center" | 168.8 | |||
| align="center" | 180.2 | |||
| align="center" | 143 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ug in 25 uL | |||
| align="center" | 4.2 | |||
| align="center" | 4.2 | |||
| align="center" | 4.5 | |||
| align="center" | 3.6 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL added | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | uL 10x buffer | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="45" | uL Lambda Exonuclease (5U/uL) | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL nfH2O | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | total | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
|} | |||
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C | 2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C |
Revision as of 18:43, 23 October 2013
BSPP Control (Started 10/22/2013)
DpnII Digestion (Started 10/22/2013
3. Column Purification, elute with 30 uL EB 4. Measure concentrations in nanodrop
Nanodrop Results
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
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
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