Daniel:Notebook/HiResChrPaint/2013-9-4

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Revision as of 00:15, 5 September 2013 by >Djacobse (→‎Lambda Exonuclease Digestion)
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BSPP Control (Started 09/03/2013)[edit]

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DpnII Digestion (Started 09/03/2013)[edit]

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

Nanodrop Results[edit]

Sample BSPP 1AB BSPP 1CD BSPP 2AB BSPP 2CD
ng/uL 127.7 130.6 110.1 109.3
ug in 30 uL 3.8 3.9 3.3 3.3
total   7.7   6.6

TBE Gel[edit]

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 BSPP 1AB BSPP 1CD BSPP 2AB BSPP 2CD
2. Run for 30 minutes at 200V
3. Stain with 3 uL SYBR gold for 2 minutes

Gel Image

File:2013-09-04-DpnIIDigest-BSPP.jpg


The gel looks fine, so I'm going to proceed as usual.

Lambda Exonuclease Digestion[edit]

1. Reaction table
Sample BSPP 1AB BSPP 1CD BSPP 2AB BSPP 2CD
ng/uL 127.7 130.6 110.1 109.3
ug in 25 uL 3.2 3.3 2.8 2.7
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
5. Measure Concentration in Nanodrop

ssNanodrop Results[edit]

ssProbe BSPP 1 BSPP 2
ng/uL 119 100.3
ug in 20 uL 2.4 2.0