Daniel:Notebook/HiResChrPaint/2013-5-13: Difference between revisions
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3. Pool samples (4x50 -> 1x200 uL) | 3. Pool samples (4x50 -> 1x200 uL) | ||
4. ssColumn Purification | 4. ssColumn Purification | ||
==TBU Gel Quantification== | |||
1. Prepared samples according to table |
Revision as of 21:37, 13 May 2013
Probe Prep Test 2 (Started 5/10/2013)
TBE Gel
1. 0.5 uL sample; 6 uL TBE, 2 uL dye per sample 2. Gel Arrangement(5-12 are digested samples that have been column purified, ready to use for lambda exo digestion):
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 25bp Ladder | V6S3A Undigested | V4S3C Pre-Column | V6S3B Pre-Column | V4S3A | V4S3B | V4S3C | V4S3D | V6S3A | V6S3B | V6S3C | V6S3D |
3. Loaded 7 uL per sample 4. Ran gel for 30 minutes at 200V 5. Stained 2 minutes with 3 uL SYBR gold
Gel Results
Lambda Exonuclease Digestion
Since I still have plenty of DNA, I'm continuing with the lambda exonuclease digestion. At the end I will be pooling the samples and running them on the ssDNA column.
1. Prepared samples according to table:
Sample | V4S3A | V4S3B | V4S3C | V4S3D | V6S3A | V6S3B | V6S3C | V6S3D |
ng/uL | 115.1 | 138.1 | 120.3 | 124.4 | 138.9 | 131.4 | 133.5 | 127.3 |
uL sample | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 |
ug sample | 2.9 | 3.5 | 3.0 | 3.1 | 3.5 | 3.3 | 3.3 | 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 1 hour at 37C followed by 10 minutes at 75C 3. Pool samples (4x50 -> 1x200 uL) 4. ssColumn Purification
TBU Gel Quantification
1. Prepared samples according to table