AlanFung:LabNotes/2014/2014-9-5: Difference between revisions
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*Remove and discard the supernatant | *Remove and discard the supernatant | ||
*Keep the tube on the magnet, add 200l of 80% ETOH | *Keep the tube on the magnet, add 200l of 80% ETOH | ||
*Incubate | *Incubate the tube at Rt for 30sec | ||
*Remove and discard the ETOH | |||
*Repeat the washing | |||
*Allow beads to dry at RT (Do not over dry beads) | |||
*Remove tubes from magnet |
Revision as of 01:03, 5 September 2014
KAPA Library Preparation Kit illumina Platforms
Parts
- Kapa Library Preparation Kit illumina platforms KR0935 V1.13
Samples
- For detailed sample information, please refer to Noi's page [[1]]
Sample | Tumor | Sample | Amount | Qubit | 100ng | Water | Condition |
ID | Type | DNA Type | ng | ng/ul | 10ng | ul | |
6-T-1 | Colon | Tumor | 100 | 43.4 | 2.30 | 47.7 | Sheared |
6-T-2 | Colon | Tumor | 100 | 31.6 | 3.16 | 46.8 | Sheared |
6-T-3 | Colon | Tumor | 100 | 31.5 | 3.17 | 46.8 | Sheared |
6-T-4 | Colon | Tumor | 100 | 41.7 | 2.40 | 47.6 | Sheared |
6-T-5 | Colon | Tumor | 100 | 30.5 | 3.28 | 46.7 | Sheared |
7-T-1 | Lung | Tumor | 100 | 30.7 | 3.26 | 46.7 | Sheared |
7-T-2 | Lung | Tumor | 100 | 44.6 | 2.24 | 47.8 | Sheared |
7-T-3 | Lung | Tumor | 100 | 35.9 | 2.79 | 47.2 | Sheared |
7-T-4 | Lung | Tumor | 100 | 33.8 | 2.96 | 47.0 | Sheared |
7-T-5 | Lung | Tumor | 100 | 20.4 | 4.90 | 45.1 | Sheared |
PC-T_1 | Pancreas | Tumor | 100 | 82.7 | 1.21 | 48.8 | Sheared |
PC-T_2 | Pancreas | Tumor | 100 | 35.8 | 2.79 | 47.2 | Sheared |
PC-T_4 | Pancreas | Tumor | 100 | 42.8 | 2.34 | 47.7 | Sheared |
PC-T_6 | Pancreas | Tumor | 100 | 15.2 | 6.58 | 43.4 | Sheared |
PC-T_7 | Pancreas | Tumor | 100 | 16.3 | 6.13 | 43.9 | Sheared |
NORM1-02 P1 | n/a | Cell-Free | 10 | 1.66 | 6.02 | 43.98 | non-Sheared |
NORM1-03 P1 | n/a | Cell-Free | 10 | 0.77 | 12.99 | 37.01 | non-Sheared |
COLO2-03 | n/a | Cell-Free | 10 | 1.11 | 9.01 | 40.99 | non-Sheared |
cfDNA | n/a | Cell-Free | 10 | 7.09 | 1.41 | 48.59 | non-Sheared |
Workflow
End Repair Reaction Setup
Component | 1 Library | 19 Libraries + 5% |
End Repair Master Mix: | ||
Water | 8 | 159.6 |
10X Kapa End Repair Buffer | 7 | 139.65 |
Kapa End Repair Enzyme Mix | 5 | 99.75 |
Total master mix volume | 20 | 399 |
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
End Repair Cleanup
- Add 120ul of diluted AMPure beads, mix by pipetting up and down
- Incubate for 5-15mins to allow the DNA to bind to beads
- Plate tubes onto a magnet to capture beads, incubate until liquid is clear
- Remove and discard the supernatant
- Keep the tube on the magnet, add 200l of 80% ETOH
- Incubate the tube at Rt for 30sec
- Remove and discard the ETOH
- Repeat the washing
- Allow beads to dry at RT (Do not over dry beads)
- Remove tubes from magnet