Daniel:Notebook/HiResChrPaint/2013-6-7: Difference between revisions

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=Probe Prep Test 2 ([[Daniel:Notebook/HiResChrPaint/2013-5-10|Started 5/10/2013]])=
=Probe Prep Test 3 ([[Daniel:Notebook/HiResChrPaint/2013-5-10|Started 5/10/2013]])=


[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]
[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]

Latest revision as of 00:32, 11 June 2013

Probe Prep Test 3 (Started 5/10/2013)[edit]

Back to Calendar

Lambda Exonuclease Digestion (Beta group, no aa-dUTP)[edit]

Going to finish lambda exonuclease digestion by taking care of the beta group.  I won't be able to go any further on this, however,
until the ULYSIS dye and the column purification are received.
Protocol:
1. Set up
Sample V4S2A V4S2B V4S2C V4S2D V6S2A V6S2B V6S2C V6S2D
ng/uL sample 126.4 122.6 108.5 112.2 129.7 112.5 140.4 128.7
uL sample 25 25 25 25 25 25 25 25
ug sample 3.2 3.1 2.7 2.8 3.2 2.8 3.5 3.2
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 nf H20 17 17 17 17 17 17 17 17
total 50 50 50 50 50 50 50 50
2. Incubate at 37C for 1 hour followed by 10 minute heat shock at 75C
3. Pool samples into single tube (4x50uL->1x200uL)
4. ssDNA column purification, elute with 10 uL nfH2O