Daniel:Notebook/HiResChrPaint/2013-7-17: Difference between revisions

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  2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C
  2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C
  3. ssDNA column purification
  3. ssDNA column purification, elute with 10 uL
  4. '''Measure Concentration in Nanodrop''' (different from before)
  4. '''Measure Concentration in Nanodrop''' (different from before)

Revision as of 19:19, 17 July 2013

S3 Probe Preparation (Started 7/16/2013)

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DpnII Digestion

3. Column Purification, elute with 30 uL EB
4. Measure concentrations in nanodrop

Nanodrop Results

Sample V4S3A V4S3B V4S3C V4S3D V6S3A V6S3B V6S3C V6S3D
ng/uL 84.9 82.3 85.4 86.4 107.6 98.2 107.4 109.2
ug in 30 uL 2.5 2.5 2.6 2.6 3.2 2.9 3.2 3.3
total ug       10.2       12.7

DpnII Digest gel

1. Lanes; (5.5 uL TBE; 2 uL 6x buffer, 0.5 uL sample; load 7 uL for all)


Lane 1 2 3 4 5 6 7 8 9 10 11 12
Sample 25bp ladder V4S3A V4S3B V4S3C V4S3D V6S3A V6S3B V6S3C V6S3D
2. Run for 25 minutes at 250V

Gel Image

File:DpnIIDigest 071713.jpg

Lambda Exonuclease Digestion

1. Reaction table
Sample V4S3A V4S3B V4S3C V4S3D V6S3A V6S3B V6S3C V6S3D
ng/uL 84.9 82.3 85.4 86.4 107.6 98.2 107.4 109.2
ug in 25 uL 2.1 2.1 2.1 2.2 2.7 2.5 2.7 2.7
uL added 25 25 25 25 25 25 25 25
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 nfH2O 17 17 17 17 17 17 17 17
total 50 50 50 50 50 50 50 50
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C
3. ssDNA column purification, elute with 10 uL
4. Measure Concentration in Nanodrop (different from before)