AlanFung:LabNotes/2014/2014-9-5: Difference between revisions
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*Kapa Library Preparation Kit illumina platforms KR0935 V1.13 | *Kapa Library Preparation Kit illumina platforms KR0935 V1.13 | ||
===Samples=== | ===Samples=== | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> | |||
|- style="background-color:#BFBFBF" align="center" | |||
| width="151" height="14" | Sample | |||
| width="65" | Amount | |||
| width="113" | Qubit ng/ul | |||
| width="65" | 10/100 ng | |||
| width="65" | Water | |||
| width="65" | Condition | |||
|- align="center" | |||
| height="14" | 6T1 | |||
| 100ng | |||
| align="center" | 43.4 | |||
|style="color:#0000FF" align="center" | 2.30 | |||
| align="center" | 47.7 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 6T2 | |||
| 100ng | |||
| align="center" | 31.6 | |||
|style="color:#0000FF" align="center" | 3.16 | |||
| align="center" | 46.8 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 6T3 | |||
| 100ng | |||
| align="center" | 31.5 | |||
|style="color:#0000FF" align="center" | 3.17 | |||
| align="center" | 46.8 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 6T4 | |||
| 100ng | |||
| align="center" | 41.7 | |||
|style="color:#0000FF" align="center" | 2.40 | |||
| align="center" | 47.6 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 6T5 | |||
| 100ng | |||
| align="center" | 30.5 | |||
|style="color:#0000FF" align="center" | 3.28 | |||
| align="center" | 46.7 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 7T1 | |||
| 100ng | |||
| align="center" | 30.7 | |||
|style="color:#0000FF" align="center" | 3.26 | |||
| align="center" | 46.7 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 7T2 | |||
| 100ng | |||
| align="center" | 44.6 | |||
|style="color:#0000FF" align="center" | 2.24 | |||
| align="center" | 47.8 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 7T3 | |||
| 100ng | |||
| align="center" | 35.9 | |||
|style="color:#0000FF" align="center" | 2.79 | |||
| align="center" | 47.2 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 7T4 | |||
| 100ng | |||
| align="center" | 33.8 | |||
|style="color:#0000FF" align="center" | 2.96 | |||
| align="center" | 47.0 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | 7T5 | |||
| 100ng | |||
| align="center" | 20.4 | |||
|style="color:#0000FF" align="center" | 4.90 | |||
| align="center" | 45.1 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | PCT1 | |||
| 100ng | |||
| align="center" | 82.7 | |||
|style="color:#0000FF" align="center" | 1.21 | |||
| align="center" | 48.8 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | PCT2 | |||
| 100ng | |||
| align="center" | 35.8 | |||
|style="color:#0000FF" align="center" | 2.79 | |||
| align="center" | 47.2 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | PCT4 | |||
| 100ng | |||
| align="center" | 42.8 | |||
|style="color:#0000FF" align="center" | 2.34 | |||
| align="center" | 47.7 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | PCT6 | |||
| 100ng | |||
| align="center" | 15.2 | |||
|style="color:#0000FF" align="center" | 6.58 | |||
| align="center" | 43.4 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | PCT7 | |||
| 100ng | |||
| align="center" | 16.3 | |||
|style="color:#0000FF" align="center" | 6.13 | |||
| align="center" | 43.9 | |||
| Sheared | |||
|- align="center" | |||
| height="14" | NORM1-02 P1 | |||
| 10ng | |||
| align="center" | 1.66 | |||
|style="color:#FF0000" align="center" | 6.02 | |||
| align="center" | 43.98 | |||
| non-Sheared | |||
|- align="center" | |||
| height="14" | NORM1-03 P1 | |||
| 10ng | |||
| align="center" | 0.77 | |||
|style="color:#FF0000" align="center" | 12.99 | |||
| align="center" | 37.01 | |||
| non-Sheared | |||
|- align="center" | |||
| height="14" | COLO2-03 | |||
| 10ng | |||
| align="center" | 1.11 | |||
|style="color:#FF0000" align="center" | 9.01 | |||
| align="center" | 40.99 | |||
| non-Sheared | |||
|- align="center" | |||
| height="14" | cfDNA | |||
| 10ng | |||
| align="center" | 7.09 | |||
|style="color:#FF0000" align="center" | 1.41 | |||
| align="center" | 48.59 | |||
| non-Sheared | |||
|} | |||
==Workflow== | ==Workflow== | ||
===End Repair Reaction Setup=== | ===End Repair Reaction Setup=== |
Revision as of 22:55, 4 September 2014
KAPA Library Preparation Kit illumina Platforms
Parts
- Kapa Library Preparation Kit illumina platforms KR0935 V1.13
Samples
Sample | Amount | Qubit ng/ul | 10/100 ng | Water | Condition |
6T1 | 100ng | 43.4 | 2.30 | 47.7 | Sheared |
6T2 | 100ng | 31.6 | 3.16 | 46.8 | Sheared |
6T3 | 100ng | 31.5 | 3.17 | 46.8 | Sheared |
6T4 | 100ng | 41.7 | 2.40 | 47.6 | Sheared |
6T5 | 100ng | 30.5 | 3.28 | 46.7 | Sheared |
7T1 | 100ng | 30.7 | 3.26 | 46.7 | Sheared |
7T2 | 100ng | 44.6 | 2.24 | 47.8 | Sheared |
7T3 | 100ng | 35.9 | 2.79 | 47.2 | Sheared |
7T4 | 100ng | 33.8 | 2.96 | 47.0 | Sheared |
7T5 | 100ng | 20.4 | 4.90 | 45.1 | Sheared |
PCT1 | 100ng | 82.7 | 1.21 | 48.8 | Sheared |
PCT2 | 100ng | 35.8 | 2.79 | 47.2 | Sheared |
PCT4 | 100ng | 42.8 | 2.34 | 47.7 | Sheared |
PCT6 | 100ng | 15.2 | 6.58 | 43.4 | Sheared |
PCT7 | 100ng | 16.3 | 6.13 | 43.9 | Sheared |
NORM1-02 P1 | 10ng | 1.66 | 6.02 | 43.98 | non-Sheared |
NORM1-03 P1 | 10ng | 0.77 | 12.99 | 37.01 | non-Sheared |
COLO2-03 | 10ng | 1.11 | 9.01 | 40.99 | non-Sheared |
cfDNA | 10ng | 7.09 | 1.41 | 48.59 | non-Sheared |
Workflow
End Repair Reaction Setup
Component | 1 Library | 15 Libraries + 5% |
End Repair Master Mix: | ||
Water | 8 | 126 |
10X Kapa End Repair Buffer | 7 | 110.25 |
Kapa End Repair Enzyme Mix | 5 | 78.75 |
Total master mix volume | 20 | 315 |
Fragmented, double-stranded DNA | 50ul |
End Repair Master Mix | 20ul |
Total rection Mix | 70ul |
- Mix, and incubate at 20C for 30mins
- Proceed immediately to the next step