Daniel:Notebook/HiResChrPaint/2013-6-7: Difference between revisions
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>Djacobse (Created page with "=Probe Prep Test 2 (Started 5/10/2013)= Back to Calendar ==Lambda Exonuclease Digestion (Beta g...") |
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2. Incubate at 37C for 1 hour followed by 10 minute heat shock at 75C | 2. Incubate at 37C for 1 hour followed by 10 minute heat shock at 75C | ||
3. Pool samples into single tube (4x50uL->1x200uL) | 3. Pool samples into single tube (4x50uL->1x200uL) | ||
4. ssDNA column purification | 4. ssDNA column purification, elute with 10 uL nfH2O |
Revision as of 23:19, 7 June 2013
Probe Prep Test 2 (Started 5/10/2013)
Lambda Exonuclease Digestion (Beta group, no aa-dUTP)
Going to finish lambda exonuclease digestion by taking care of the beta group. I won't be able to go any further on this, however, until the ULYSIS dye and the column purification are received.
Protocol: 1. Set up
Sample | V4S2A | V4S2B | V4S2C | V4S2D | V6S2A | V6S2B | V6S2C | V6S2D |
ng/uL sample | 126.4 | 122.6 | 108.5 | 112.2 | 129.7 | 112.5 | 140.4 | 128.7 |
uL sample | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 |
ug sample | 3.2 | 3.1 | 2.7 | 2.8 | 3.2 | 2.8 | 3.5 | 3.2 |
uL 10x buffer | 5 | 5 | 5 | 5 | 5 | 5 | 5 | 5 |
uL lambda exonuclease (5U/uL) | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
uL nf H20 | 17 | 17 | 17 | 17 | 17 | 17 | 17 | 17 |
total | 50 | 50 | 50 | 50 | 50 | 50 | 50 | 50 |
2. Incubate at 37C for 1 hour followed by 10 minute heat shock at 75C 3. Pool samples into single tube (4x50uL->1x200uL) 4. ssDNA column purification, elute with 10 uL nfH2O