Daniel:Notebook/HiResChrPaint/2014-7-9: Difference between revisions
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Scary that both of the columns yielded the same percent (I double checked the math). As a note, taking into account that the theoretical maximum yield is 80% (since we remove 20bp out of 100 on each strand), this is approximately a '''90% yield''' from the column. | Scary that both of the columns yielded the same percent (I double checked the math). As a note, taking into account that the theoretical maximum yield is 80% (since we remove 20bp out of 100 on each strand), this is approximately a '''90% yield''' from the column. | ||
===TBE Gel=== | |||
'''Gel Image''' | |||
[[Image:2014_07_09_DpnII.jpg|500px]] | |||
==Lambda Exonuclease Digestion== | ==Lambda Exonuclease Digestion== |
Revision as of 21:26, 9 July 2014
Strand Control (Started 07/08/2014)
DpnII Digest (from 07/08/2014)
- Column Purification, elute with 40 uL
- Measure concentration in nanodrop
Nanodrop Results
Sample | V6S2AB | V6S2CD | V6S3AB | V6S3CD |
ng/uL | 122.4 | 110.7 | 137 | 133.5 |
ug in 40 uL | 4.9 | 4.4 | 5.5 | 5.3 |
ug total | 9.3 | 10.8 | ||
Yield (Max ~80%) | 71.18 | 71.18 |
Scary that both of the columns yielded the same percent (I double checked the math). As a note, taking into account that the theoretical maximum yield is 80% (since we remove 20bp out of 100 on each strand), this is approximately a 90% yield from the column.
TBE Gel
Gel Image
Lambda Exonuclease Digestion
- Reaction Table
Sample | V6S2AB | V6S2CD | V6S3AB | V6S3CD |
ng/uL | 122.4 | 110.7 | 137 | 133.5 |
ug in 35 uL | 4.3 | 3.9 | 4.8 | 4.7 |
uL sample | 35 | 35 | 35 | 35 |
uL 10X buffer | 5 | 5 | 5 | 5 |
uL Lambda Exonuclease (5 U/uL) | 3 | 3 | 3 | 3 |
uL nfH2O | 7 | 7 | 7 | 7 |
Total | 50 | 50 | 50 | 50 |
- Incubate for 1 hr at 37C
- Heat inactivate for 10 minutes at 75C
- Purify with Zymo column, elute with 20 uL nfH2O