Sam:LabNotes/Microbiome-new/2010-11-17: Difference between revisions
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>Sam Chiang (Created page with '='''MDA samples preparation for Covaris shearing-(single E.coli amplicons)'''= ==Background== *I want attempt to prepare more sheared E.coli SAGs (from short-vs-long ampificatio…') |
>Sam Chiang No edit summary |
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===Procedures=== | ===Procedures=== | ||
*Label the Covaris microtubes. | *Label the Covaris microtubes. Open the caps and leave tubes on 96-well rack. | ||
*UV-treat the tubes and pipettes for 15min. | |||
**Thaw the tRNA carrier (stored in -80C). Use the "2"-tRNA(96ng/uL) | **Thaw the tRNA carrier (stored in -80C). Use the "2"-tRNA(96ng/uL) | ||
Sample no. Sample name Conc.(ng/uL) Sample(uL) 330ng-tRNA("2") H2O Total DNA(ng) | Sample no. Sample name Conc.(ng/uL) Sample(uL) 330ng-tRNA("2") H2O Total DNA(ng) | ||
--------------------------------------------------------------------------------------------------- | --------------------------------------------------------------------------------------------------- | ||
HC sam080610_2-1_2h 0.685 100 3.43 0 68.5ng | |||
2-S sam080610_2-2_2h 0.703 100 3.43 0 70.3ng | 2-S sam080610_2-2_2h 0.703 100 3.43 0 70.3ng | ||
3-S sam081210_1-5_2h 0.704 100 3.43 0 70.4ng | 3-S sam081210_1-5_2h 0.704 100 3.43 0 70.4ng |
Revision as of 07:28, 18 November 2010
MDA samples preparation for Covaris shearing-(single E.coli amplicons)
Background
- I want attempt to prepare more sheared E.coli SAGs (from short-vs-long ampification) for Illumina library construction.
- I didn't do any additional amplification on the short-amplified E.coli SAGs since the over-dose MDA reaction buffer may interfere the library construction.
QUBIT Sample quantification
Procedures
- Label the Covaris microtubes. Open the caps and leave tubes on 96-well rack.
- UV-treat the tubes and pipettes for 15min.
- Thaw the tRNA carrier (stored in -80C). Use the "2"-tRNA(96ng/uL)
Sample no. Sample name Conc.(ng/uL) Sample(uL) 330ng-tRNA("2") H2O Total DNA(ng) --------------------------------------------------------------------------------------------------- HC sam080610_2-1_2h 0.685 100 3.43 0 68.5ng 2-S sam080610_2-2_2h 0.703 100 3.43 0 70.3ng 3-S sam081210_1-5_2h 0.704 100 3.43 0 70.4ng 4-S sam081210_2-5_2h 0.693 100 3.43 0 69.3ng 1-L sam080610_2-1_10h 220.0 2.27 - 97.7 500.0ng 2-L sam080610_2-2_10h 322.0 1.55 - 98.5 500.0ng 3-L sam081210_1-5_10h 540.0 0.92 - 99.0 500.0ng 4-L sam081210_2-5_10h 551.0 0.90 - 99.0 500.0ng
- 100uL sample suspension were carefully transferred into microtubes.
- Samples tubes were tapped on the rack and packed in a paper box with three disposable cooler.
- The package was shipped to the Sara through FedEx(track# 4490-5094-2339 on 10-19-'10).