Daniel:Notebook/HiResChrPaint/2013-6-21: Difference between revisions
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>Djacobse (Created page with "=S1 Probe Preparation (Started 6/20/2013)= Back to Calendar ==DpnII Digestion (Continued from [...") |
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Revision as of 20:39, 21 June 2013
S1 Probe Preparation (Started 6/20/2013)
DpnII Digestion (Continued from 6/20/2013)
3. Pooled all V4 samples (4->1) and 2 V6 samples (4->2) 4. Column purification, elute with 40 uL EB 5. Measure Concentration in Nanodrop 6. TBE gel to ensure proper digestion
Nanodrop Results
Sample | V4S1 | V6S1AB | V6S1CD |
ng/uL | 66.7 | 118.3 | 120.6 |
ug in 40 uL | 2.7 | 4.7 | 4.8 |
TBE gel
1. Prepared samples 5.5 uL TBE, 0.5 uL sample, 2 uL 6x dye 2. Sample matrix
Lane | 4 | 5 | 6 | 7 | 8 | 9 |
Sample | 25 bp ladder | V4S1B Undigested | V6S1B Undigested | V4S1 | V6S1AB | V6S1CD |
3. Ran gel for 20 minutes at 220 V, stained for 2 minutes with 3 uL SYBR gold
Gel Image
[[Image:]]
Lambda Exonuclease digestion
1. Reaction table
Sample | V4S1 | V6S1AB | V6S1CD |
ng/uL | 66.7 | 118.3 | 120.6 |
ug in 40 uL | 2.7 | 4.7 | 4.8 |
uL 10x buffer | 5 | 5 | 5 |
uL Lambda Exonuclease | 3 | 5 | 5 |
uL nfH2O | 7 | 5 | 5 |
total | 50 | 50 | 50 |
3. Incubate at 37C for 1 hour followed by 10 minute heat shock at 75C 4. ssDNA column purification, elute with 10 uL