Daniel:Notebook/HiResChrPaint/2013-5-3
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Probe Prep Test (Started 4/22/2013)[edit]
Lambda Exonuclease Test[edit]
1. Prepare samples according to following table *Enzyme 0050909; Buffer 001907;
Sample | ng/uL | uL | ng digested | uL lambda exo (5 U/uL) | uL 10X buffer | uL nf H20 | Total | Digest Time |
V4S2A | 39.6 | 8 | 316.8 | 0.5 | 5 | 36.5 | 50 | 30 min |
V4S2B | 39.6 | 8 | 316.8 | 0.5 | 5 | 36.5 | 50 | 1 hr |
V4S2C | 39.6 | 8 | 316.8 | 0.5 | 5 | 36.5 | 50 | 2 hr |
V6S2A | 30.6 | 8 | 244.8 | 0.5 | 5 | 36.5 | 50 | 30 min |
V6S2B | 30.6 | 8 | 244.8 | 0.5 | 5 | 36.5 | 50 | 1 hr |
V6S2C | 30.6 | 8 | 244.8 | 0.5 | 5 | 36.5 | 50 | 2 hr |
2. After each incubation include a 10 minute heat shock at 75 C 3. Mix 45 uL sample with 250 uL nf H20 in a 1.5 mL eppendorf 4. Ethanol Precipitation
Ethanol Precipitation[edit]
1. 3X 100% EtOH; 0.1X 3 M NaOAc; 1/300 glycoblue 2. Incubate overnight at -80 C (continued 5/4/2013)