Daniel:Notebook/HiResChrPaint/2013-6-6

From ZhangLabWiki
Revision as of 21:45, 6 June 2013 by >Djacobse
Jump to navigation Jump to search

Probe Prep 3 (Started 5/29/2013)

Back to Calendar

Summary of DpnII Digestion Reactions

Alpha Group (aa-dUTP present)

Sample V4S2A V4S2B V4S2C V4S2D V6S2A V6S2B V6S2C V6S2D
ng/uL 121.5 127.1 119.1 121.8 136.7 122.4 119.2 147.7
ug left 3.6 3.8 3.6 3.7 4.1 3.7 3.6 4.4
total 14.7 15.8

Beta Group (no aa-dUTP)

Sample V4S2A V4S2B V4S2C V4S2D V6S2A V6S2B V6S2C V6S2D
ng/uL 126.4 122.6 108.5 112.2 129.7 112.5 140.4 128.7
ug left 3.8 3.7 3.3 3.4 3.9 3.4 4.2 3.9
total 14.1 15.3

Lambda Exonuclease Digestion (Alpha Group)

1. Prepare samples
Sample V4S2A V4S2B V4S2C V4S2D V6S2A V6S2B V6S2C V6S2D
ng/uL sample 121.5 127.1 119.1 121.8 136.7 122.4 119.2 147.7
uL sample 25 25 25 25 25 25 25 25
ug sample 3.0375 3.1775 2.9775 3.045 3.4175 3.06 2.98 3.6925
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