Daniel:Notebook/HiResChrPaint/2013-6-21: Difference between revisions

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  1. Reaction table
  1. Reaction table
2. {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
 
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="font-size:12pt;font-weight:bold" align="center"
|- style="font-size:12pt;font-weight:bold" align="center"
| width="96" height="15" | Sample
| width="96" height="15" | Sample

Revision as of 20:39, 21 June 2013

S1 Probe Preparation (Started 6/20/2013)

Back to Calendar

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