Daniel:Notebook/HiResChrPaint/2014-7-9: Difference between revisions

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>Djacobse
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#Reaction Table
#Reaction Table


{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="background-color:#C5D9F1;font-size:12pt;font-weight:bold"
| width="119" height="15"  valign="bottom" | Sample
| width="65" align="center" valign="bottom" | V6S2AB
| width="65" align="center" valign="bottom" | V6S2CD
| width="65" align="center" valign="bottom" | V6S3AB
| width="65" align="center" valign="bottom" | V6S3CD
|- style="font-size:12pt"
|style="font-weight:bold" height="15"  valign="bottom" | ng/uL
| align="center" align="center" valign="bottom" | 122.4
| align="center" align="center" valign="bottom" | 110.7
| align="center" align="center" valign="bottom" | 137
| align="center" align="center" valign="bottom" | 133.5
|- style="background-color:#D9D9D9;font-size:12pt"
|style="font-weight:bold" height="15"  valign="bottom" | ug in 35 uL
| align="center" align="center" valign="bottom" | 4.3
| align="center" align="center" valign="bottom" | 3.9
| align="center" align="center" valign="bottom" | 4.8
| align="center" align="center" valign="bottom" | 4.7
|- style="font-size:12pt"
|style="font-weight:bold" height="15"  valign="bottom" | uL sample
| align="center" align="center" valign="bottom" | 35
| align="center" align="center" valign="bottom" | 35
| align="center" align="center" valign="bottom" | 35
| align="center" align="center" valign="bottom" | 35
|- style="background-color:#D9D9D9;font-size:12pt"
|style="font-weight:bold" height="15"  valign="bottom" | uL 10X buffer
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5
|- style="font-size:12pt"
|style="font-weight:bold" height="30"  valign="bottom" | uL Lambda Exonuclease (5 U/uL)
| align="center" align="center" valign="bottom" | 3
| align="center" align="center" valign="bottom" | 3
| align="center" align="center" valign="bottom" | 3
| align="center" align="center" valign="bottom" | 3
|- style="background-color:#D9D9D9;font-size:12pt"
|style="font-weight:bold" height="15"  valign="bottom" | uL nfH2O
| align="center" align="center" valign="bottom" | 7
| align="center" align="center" valign="bottom" | 7
| align="center" align="center" valign="bottom" | 7
| align="center" align="center" valign="bottom" | 7
|- style="font-size:12pt"
|style="font-weight:bold" height="15"  valign="bottom" | Total
| align="center" align="center" valign="bottom" | 50
| align="center" align="center" valign="bottom" | 50
| align="center" align="center" valign="bottom" | 50
| align="center" align="center" valign="bottom" | 50
|}





Revision as of 20:16, 9 July 2014

Strand Control (Started 07/08/2014)

Back to Calendar

DpnII Digest (from 07/08/2014)

  1. Column Purification, elute with 40 uL
  2. 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.

Lambda Exonuclease Digestion

  1. 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


  1. Incubate for 1 hr at 37C
  2. Heat inactivate for 10 minutes at 75C
  3. Purify with Zymo column, elute with 20 uL nfH2O