Daniel:Notebook/HiResChrPaint/2013-4-27

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Revision as of 23:31, 27 April 2013 by >Djacobse
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Probe Prep Test (Started 4/22/2013)

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Ethanol Precipitation (Continued from 4/26/2013)

5. Took samples from -80 and centrifuged at 14000 rpm for 25 minutes at 4 C
6. Removed supernatant and added 750 uL 75% EtOH
7. Centrifugd at 14000 rpm for 15 minutes at 4 C
8. Removed most supernatant with pipette and then dried in hood
9. Resuspended in 8 uL nuclease free water

TBU gel Quantification

1. Preheated TBE buffer in 60 C incubator
2. microwaved for 1 minute
3. Added hot TBE to gel box and immediately preran gel for 20 minutes at 250V
4. Created samples according to following table
Lane 3 4 5 6 7 8 9 10
Sample Ladder Ladder V4S3 V4S3 V6S3 V6S3 Ladder Buffer
uL sample 0.5 1 0.5 0.5 0.5 0.5 0.5 0
uL hot TBE 5.5 5 5.5 5.5 5.5 5.5 5.5 6
uL 2x buffer 6 6 6 6 6 6 6 6
loaded 12 12 12 12 12 12 12 12
5. Incubated samples at 65C for 8 minutes
6. Cold snapped with ice box
7. Loaded 12 uL each sample and ran gel for 20 minutes at 250V
8. Stained with 3 uL SYBR gold for 25 minutes