Daniel:Notebook/HiResChrPaint/2013-3-27: Difference between revisions
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>Djacobse (Created page with "=Probe Set Prep (Started 3/26/2013)= Back to Calendar ==Expansion PCR and Ethanol Precipitation==") |
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[[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | ||
== | ==Ethanol Precipitation ([[Daniel:Notebook/HiResChrPaint/2013-3-26|From 3/26/2013]])== | ||
4. Centrifuge at 3000 rpm for 30 min, 4 C | |||
5. Remove Supernatant and add 600 uL chilled 75% EtOH | |||
6. Transfer DNA pellet to a 1.5 mL tube | |||
7. Spin at 14000 rpm for 5 min | |||
8. Dry the pellet in the hood and resuspend with 100 uL ddH2O | |||
9. Run 2 [[Daniel:Protocols/Qiaquick_Column|Qiaquick columns]] per sample, elute with 40 uL for each column | |||
10. Pool samples (16 columns -> 8 samples) | |||
11. Measure concentration with nanodrop | |||
{| 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="16" | Probe | |||
| width="65" | ng/uL | |||
| width="65" | uL | |||
| width="65" | ng | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V4S1 | |||
| align="center" | 36.4 | |||
| align="center" | 80 | |||
| align="center" | 2912 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V4S2 | |||
| align="center" | 139.7 | |||
| align="center" | 80 | |||
| align="center" | 11176 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V4S3 | |||
| align="center" | 76.2 | |||
| align="center" | 80 | |||
| align="center" | 6096 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V4S4 | |||
| align="center" | 47.8 | |||
| align="center" | 80 | |||
| align="center" | 3824 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V6S1 | |||
| align="center" | 84.4 | |||
| align="center" | 80 | |||
| align="center" | 6752 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V6S2 | |||
| align="center" | 111.9 | |||
| align="center" | 80 | |||
| align="center" | 8952 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V6S3 | |||
| align="center" | 131.4 | |||
| align="center" | 80 | |||
| align="center" | 10512 | |||
|- style="font-size:12pt" align="center" | |||
| height="16" | V6S4 | |||
| align="center" | 106.9 | |||
| align="center" | 80 | |||
| align="center" | 8552 | |||
|} |
Latest revision as of 23:44, 17 April 2013
Probe Set Prep (Started 3/26/2013)[edit]
Ethanol Precipitation (From 3/26/2013)[edit]
4. Centrifuge at 3000 rpm for 30 min, 4 C 5. Remove Supernatant and add 600 uL chilled 75% EtOH 6. Transfer DNA pellet to a 1.5 mL tube 7. Spin at 14000 rpm for 5 min 8. Dry the pellet in the hood and resuspend with 100 uL ddH2O 9. Run 2 Qiaquick columns per sample, elute with 40 uL for each column 10. Pool samples (16 columns -> 8 samples) 11. Measure concentration with nanodrop
Probe | ng/uL | uL | ng |
V4S1 | 36.4 | 80 | 2912 |
V4S2 | 139.7 | 80 | 11176 |
V4S3 | 76.2 | 80 | 6096 |
V4S4 | 47.8 | 80 | 3824 |
V6S1 | 84.4 | 80 | 6752 |
V6S2 | 111.9 | 80 | 8952 |
V6S3 | 131.4 | 80 | 10512 |
V6S4 | 106.9 | 80 | 8552 |