Daniel:Notebook/HiResChrPaint/2013-5-3

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Probe Prep Test (Started 4/22/2013)[edit]

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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)