Daniel:Notebook/HiResChrPaint/2013-11-16: Difference between revisions
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>Djacobse (Created page with "=Positive Control 2 (V6S3 & V6S4, Started 11/15/2013)= Back to Calendar ==Lambda Exonuclease D...") |
>Djacobse |
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|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="15" | ng/uL | |style="font-weight:bold" height="15" | ng/uL | ||
| align=" | | align="center" | 169.9 | ||
| align=" | | align="center" | 180 | ||
| align=" | | align="center" | 169.4 | ||
| align=" | | align="center" | 180.1 | ||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="15" | ug in 25 uL | |style="font-weight:bold" height="15" | ug in 25 uL | ||
| align=" | | align="center" | 4.2 | ||
| align=" | | align="center" | 4.5 | ||
| align=" | | align="center" | 4.2 | ||
| align=" | | align="center" | 4.5 | ||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="15" | uL added | |style="font-weight:bold" height="15" | uL added | ||
| align=" | | align="center" | 25 | ||
| align=" | | align="center" | 25 | ||
| align=" | | align="center" | 25 | ||
| align=" | | align="center" | 25 | ||
| | |||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="30" | uL 10x buffer | |style="font-weight:bold" height="30" | uL 10x buffer | ||
| align=" | | align="center" | 5 | ||
| align=" | | align="center" | 5 | ||
| align=" | | align="center" | 5 | ||
| align=" | | align="center" | 5 | ||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="45" | uL Lambda Exonuclease (5U/uL) | |style="font-weight:bold" height="45" | uL Lambda Exonuclease (5U/uL) | ||
| align=" | | align="center" | 3 | ||
| align=" | | align="center" | 3 | ||
| align=" | | align="center" | 3 | ||
| align=" | | align="center" | 3 | ||
| | |||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="15" | uL nfH2O | |style="font-weight:bold" height="15" | uL nfH2O | ||
| align=" | | align="center" | 17 | ||
| align=" | | align="center" | 17 | ||
| align=" | | align="center" | 17 | ||
| align=" | | align="center" | 17 | ||
|- style="font-size:12pt" align="center" | |- style="font-size:12pt" align="center" | ||
|style="font-weight:bold" height="15" | total | |style="font-weight:bold" height="15" | total | ||
| align=" | | align="center" | 50 | ||
| align=" | | align="center" | 50 | ||
| align=" | | align="center" | 50 | ||
| align=" | | align="center" | 50 | ||
|} | |} | ||
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4. [[Daniel:Protocols/ssDNA Column|ssDNA column]] purification, elute with 20 uL nfH2O | 4. [[Daniel:Protocols/ssDNA Column|ssDNA column]] purification, elute with 20 uL nfH2O | ||
5. Measure Concentration in Nanodrop | 5. Measure Concentration in Nanodrop | ||
===ssNanodrop Results=== | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" align="center" | |||
| width="65" height="15" | Sample | |||
| width="65" | ng/uL | |||
| width="65" | ug in 20 uL | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | V6S3 | |||
| align="center" | 144.3 | |||
| align="center" | 2.9 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | V6S4 | |||
| align="center" | 137.2 | |||
| align="center" | 2.7 | |||
|} |
Latest revision as of 01:41, 17 November 2013
Positive Control 2 (V6S3 & V6S4, Started 11/15/2013)[edit]
Lambda Exonuclease Digestion[edit]
1. Reaction table
Sample | V6S3AB | V6S3CD | V6S4AB | V6S4CD | |
ng/uL | 169.9 | 180 | 169.4 | 180.1 | |
ug in 25 uL | 4.2 | 4.5 | 4.2 | 4.5 | |
uL added | 25 | 25 | 25 | 25 | |
uL 10x buffer | 5 | 5 | 5 | 5 | |
uL Lambda Exonuclease (5U/uL) | 3 | 3 | 3 | 3 | |
uL nfH2O | 17 | 17 | 17 | 17 | |
total | 50 | 50 | 50 | 50 |
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C 3. Pool samples 4. ssDNA column purification, elute with 20 uL nfH2O 5. Measure Concentration in Nanodrop
ssNanodrop Results[edit]
Sample | ng/uL | ug in 20 uL |
V6S3 | 144.3 | 2.9 |
V6S4 | 137.2 | 2.7 |