Daniel:Notebook/HiResChrPaint/2013-10-23

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Revision as of 19:18, 23 October 2013 by >Djacobse (→‎TBE Gel)
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BSPP Control (Started 10/22/2013)

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DpnII Digestion (Started 10/22/2013

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

Nanodrop Results

Sample BSPP 1AB BSPP 1CD BSPP 2AB BSPP 2CD
ng/uL 166.5 168.8 180.2 143
ug in 30 uL 5.0 5.1 5.4 4.3
total   10.1   9.7

TBE Gel

1. Lanes; (5.5 uL TBE; 2 uL 6x buffer, 0.5 uL sample; load 7.2 uL for all)


Lane 1 2 3 4 5 6 7 8 9 10 11 12
Sample 10bp ladder BSPP 1AB BSPP 1CD BSPP 2AB BSPP 2CD
2. Run for 23 minutes at 220V
3. Stain with 3 uL SYBR gold for 2 minutes

Gel Image

File:20131023 DpnIIDigest.jpg

Lambda Exonuclease Digestion

1. Reaction table
Sample BSPP 1AB BSPP 1CD BSPP 2AB BSPP 2CD
ng/uL 166.5 168.8 180.2 143
ug in 25 uL 4.2 4.2 4.5 3.6
uL added 25 25 25 25
uL 10x buffer 5 5 5 5
uL Lambda Exonuclease (5U/uL) 3 3 3 3
uL nfH2O 17 17 17 17
total 50 50 50 50
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C
3. Pool samples
4. ssDNA column purification, elute with 20 uL nfH2O
5. Measure Concentration in Nanodrop

ssNanodrop Results