Daniel:Notebook/HiResChrPaint/2013-4-26: Difference between revisions
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>Djacobse (Created page with "=Probe Prep Test (Started 4/22/2013)= Back to calendar ==Lambda Exonuclease Digest==") |
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==Lambda Exonuclease Digest== | ==Lambda Exonuclease Digest== | ||
1. Prepared samples according to following table | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" align="center" | |||
| width="65" height="33" | Sample | |||
| width="65" | ng/uL | |||
| width="65" | uL sample | |||
| width="65" | uL 10x buffer | |||
| width="65" | uL Lambda Exo | |||
| width="65" | uL ddH20 | |||
| width="65" | Total | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V4S3A | |||
| align="center" | 102.2 | |||
| align="center" | 30 | |||
| align="center" | 5 | |||
| align="center" | 2 | |||
| align="center" | 13 | |||
| align="center" | 50 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V4S3B | |||
| align="center" | 86.8 | |||
| align="center" | 30 | |||
| align="center" | 5 | |||
| align="center" | 2 | |||
| align="center" | 13 | |||
| align="center" | 50 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V6S3A | |||
| align="center" | 80.1 | |||
| align="center" | 30 | |||
| align="center" | 5 | |||
| align="center" | 2 | |||
| align="center" | 13 | |||
| align="center" | 50 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V6S3B | |||
| align="center" | 77.9 | |||
| align="center" | 30 | |||
| align="center" | 5 | |||
| align="center" | 2 | |||
| align="center" | 13 | |||
| align="center" | 50 | |||
|} | |||
2. Incubate for 1 hour at 37 C | |||
3. Heat shock 75 C 10 minutes | |||
4. Ethanol Precipitation | |||
==Ethanol Precipitation== | |||
1. Pooled samples A and B (total 100 uL) | |||
2. Add 200 uL nuclease free H20 to mixture (total 300 uL) | |||
3. Add 900 uL 100% EtOH, 30 uL NaOAc, 2 uL glycoblue (2/300X) | |||
4. Incubate overnight at -80C; Continued on [[Daniel:Notebook/HiResChrPaint/2013-4-27|4/27/2013]] |
Latest revision as of 23:40, 27 April 2013
Probe Prep Test (Started 4/22/2013)[edit]
Lambda Exonuclease Digest[edit]
1. Prepared samples according to following table
Sample | ng/uL | uL sample | uL 10x buffer | uL Lambda Exo | uL ddH20 | Total |
V4S3A | 102.2 | 30 | 5 | 2 | 13 | 50 |
V4S3B | 86.8 | 30 | 5 | 2 | 13 | 50 |
V6S3A | 80.1 | 30 | 5 | 2 | 13 | 50 |
V6S3B | 77.9 | 30 | 5 | 2 | 13 | 50 |
2. Incubate for 1 hour at 37 C 3. Heat shock 75 C 10 minutes 4. Ethanol Precipitation
Ethanol Precipitation[edit]
1. Pooled samples A and B (total 100 uL) 2. Add 200 uL nuclease free H20 to mixture (total 300 uL) 3. Add 900 uL 100% EtOH, 30 uL NaOAc, 2 uL glycoblue (2/300X) 4. Incubate overnight at -80C; Continued on 4/27/2013