Daniel:Notebook/HiResChrPaint/2013-6-6: Difference between revisions
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==Lambda Exonuclease Digestion (Alpha Group)== | |||
1. Prepare samples | |||
{| 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="134" height="15" | Sample | |||
| width="65" | V4S2A | |||
| width="65" | V4S2B | |||
| width="65" | V4S2C | |||
| width="65" | V4S2D | |||
| width="65" | V6S2A | |||
| width="65" | V6S2B | |||
| width="65" | V6S2C | |||
| width="65" | V6S2D | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ng/uL sample | |||
| align="center" | 121.5 | |||
| align="center" | 127.1 | |||
| align="center" | 119.1 | |||
| align="center" | 121.8 | |||
| align="center" | 136.7 | |||
| align="center" | 122.4 | |||
| align="center" | 119.2 | |||
| align="center" | 147.7 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL sample | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
| align="center" | 25 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ug sample | |||
| align="center" | 3.0375 | |||
| align="center" | 3.1775 | |||
| align="center" | 2.9775 | |||
| align="center" | 3.045 | |||
| align="center" | 3.4175 | |||
| align="center" | 3.06 | |||
| align="center" | 2.98 | |||
| align="center" | 3.6925 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL 10x buffer | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
| align="center" | 5 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="30" | uL lambda exonuclease (5U/uL) | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
| align="center" | 3 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL nf H20 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
| align="center" | 17 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | total | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
| align="center" | 50 | |||
|} | |||
2. Incubate at 37C for 1 hour followed by 10 minute heat shock at 75C | |||
3. Pool samples into single tube (4x50uL->1x200uL) | |||
4. ssDNA column purification, elute with 10 uL nfH2O. |
Latest revision as of 23:19, 7 June 2013
Probe Prep 3 (Started 5/29/2013)[edit]
Summary of DpnII Digestion Reactions[edit]
Alpha Group (aa-dUTP present)[edit]
Sample | V4S2A | V4S2B | V4S2C | V4S2D | V6S2A | V6S2B | V6S2C | V6S2D |
ng/uL | 121.5 | 127.1 | 119.1 | 121.8 | 136.7 | 122.4 | 119.2 | 147.7 |
ug left | 3.6 | 3.8 | 3.6 | 3.7 | 4.1 | 3.7 | 3.6 | 4.4 |
total | 14.7 | 15.8 |
Beta Group (no aa-dUTP)[edit]
Sample | V4S2A | V4S2B | V4S2C | V4S2D | V6S2A | V6S2B | V6S2C | V6S2D |
ng/uL | 126.4 | 122.6 | 108.5 | 112.2 | 129.7 | 112.5 | 140.4 | 128.7 |
ug left | 3.8 | 3.7 | 3.3 | 3.4 | 3.9 | 3.4 | 4.2 | 3.9 |
total | 14.1 | 15.3 |
Lambda Exonuclease Digestion (Alpha Group)[edit]
1. Prepare samples
Sample | V4S2A | V4S2B | V4S2C | V4S2D | V6S2A | V6S2B | V6S2C | V6S2D |
ng/uL sample | 121.5 | 127.1 | 119.1 | 121.8 | 136.7 | 122.4 | 119.2 | 147.7 |
uL sample | 25 | 25 | 25 | 25 | 25 | 25 | 25 | 25 |
ug sample | 3.0375 | 3.1775 | 2.9775 | 3.045 | 3.4175 | 3.06 | 2.98 | 3.6925 |
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 at 37C for 1 hour followed by 10 minute heat shock at 75C 3. Pool samples into single tube (4x50uL->1x200uL) 4. ssDNA column purification, elute with 10 uL nfH2O.