AlanFung:LabNotes/Capturing/2012-8-10: Difference between revisions
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>Alan6017518 (Created page with "==Capturing Remaining Gimelbrant lab Sample== *Add 2 drops of mineral oil to prevent evaporation {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA...") |
>Alan6017518 |
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| align="center" | 10.3 | | align="center" | 10.3 | ||
| align="center" | 20 | | align="center" | 20 | ||
|} | |||
*Make sure we have enough reagent before start, | |||
*need 50ul probes, check | |||
*need 66ul 10X Ampligase Buffer, | |||
* | |||
===Prepare SLN Mix=== | |||
*Dilute 1mM dNTP 1 in 100 to 10uM dNTP | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> | |||
|- style="background-color:#B1A0C7;font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold;text-decoration:underline" width="270" height="13" | SLN Mix for 20ul Rxn | |||
|style="font-weight:bold;text-decoration:none" width="121" | 1x | |||
| width="41" | 16.5x | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | dNTP (10uM) | |||
| align="center" | 0.5 | |||
| align="center" | 8.25 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | Stoffel (10U/ul) | |||
| align="center" | 0.2 | |||
| align="center" | 3.30 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | Ampligase (5U/ul) | |||
| align="center" | 0.2 | |||
| align="center" | 3.30 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | H2O | |||
| align="center" | 0.1 | |||
| align="center" | 1.65 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
| align="center" height="13" | | |||
| align="center" | | |||
| align="center" | | |||
|- style="background-color:#B1A0C7;font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold;text-decoration:underline" height="13" | SLN Mix for 50ul Rxn | |||
|style="font-weight:bold;text-decoration:none" | 1x | |||
| 19.8x | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | dNTP (10uM) | |||
| align="center" | 1.25 | |||
| align="center" | 24.75 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | Stoffel (10U/ul) | |||
| align="center" | 0.5 | |||
| align="center" | 9.90 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | Ampligase (5U/ul) | |||
| align="center" | 0.5 | |||
| align="center" | 9.90 | |||
|- style="font-size:8pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="13" | H2O | |||
| align="center" | 0.25 | |||
| align="center" | 4.95 | |||
|} | |} |
Revision as of 00:29, 11 August 2012
Capturing Remaining Gimelbrant lab Sample
- Add 2 drops of mineral oil to prevent evaporation
ChIPK36me3 | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |
GM12878 | 0.90 | 40.00 | 0.45 | 2 | 7.6 | 50 |
P10 | 1.20 | 40.00 | 0.60 | 2 | 7.4 | 50 |
DF2 | 0.70 | 40.00 | 0.35 | 2 | 7.7 | 50 |
H0 | 1.00 | 40.00 | 0.50 | 2 | 7.5 | 50 |
H7 | 0.60 | 40.00 | 0.30 | 2 | 7.7 | 50 |
H14 | 0.50 | 40.00 | 0.25 | 2 | 7.8 | 50 |
H16 | 0.60 | 40.00 | 0.30 | 2 | 7.7 | 50 |
ChIPK27me3 | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |
GM12878 | 1.20 | 40.00 | 0.60 | 2 | 7.4 | 50 |
GM12878 Round 2 | 2.86 | 35.00 | 1.25 | 2 | 11.8 | 50 |
P10 | 1.30 | 40.00 | 0.65 | 2 | 7.4 | 50 |
DF2 | 2 | 40.00 | 1.00 | 2 | 7.0 | 50 |
H0 | 2.4 | 40.00 | 1.20 | 2 | 6.8 | 50 |
H7 | 1.2 | 40.00 | 0.60 | 2 | 7.4 | 50 |
H14 | 0.5 | 40.00 | 0.25 | 2 | 7.8 | 50 |
H16 | 1.3 | 40.00 | 0.65 | 2 | 7.4 | 50 |
WCE | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |
GM12878 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
GM12878 Round 2 | 2.86 | 35.00 | 1.25 | 2 | 11.8 | 50 |
P10 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
P10 Round 2 | 2.86 | 35.00 | 1.25 | 2 | 11.8 | 50 |
DF2 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
DF2 Round 2 | 2.86 | 35.00 | 1.25 | 2 | 11.8 | 50 |
H0 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
H7 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
H14 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
H16 | 35.39 | 5.65 | 2.49 | 2 | 9.9 | 20 |
cDNA | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |
GM | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
H0 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
H7 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
H14 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
H16 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
gDNA | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |
H7 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
H14 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
H16 | 38.12 | 5.25 | 2.49 | 2 | 10.3 | 20 |
- Make sure we have enough reagent before start,
- need 50ul probes, check
- need 66ul 10X Ampligase Buffer,
Prepare SLN Mix
- Dilute 1mM dNTP 1 in 100 to 10uM dNTP
SLN Mix for 20ul Rxn | 1x | 16.5x |
dNTP (10uM) | 0.5 | 8.25 |
Stoffel (10U/ul) | 0.2 | 3.30 |
Ampligase (5U/ul) | 0.2 | 3.30 |
H2O | 0.1 | 1.65 |
SLN Mix for 50ul Rxn | 1x | 19.8x |
dNTP (10uM) | 1.25 | 24.75 |
Stoffel (10U/ul) | 0.5 | 9.90 |
Ampligase (5U/ul) | 0.5 | 9.90 |
H2O | 0.25 | 4.95 |