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
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==Sample preparation for shipping (10-19-'10)==
*The samples were not shipped until we got the Covaris microtubes(cat# 520045)
**In the future, samples will be transferred to microtubes before shearing, which can help Sarah to save lots of work.
*Eight samples, in total, were prepared as following arrangements.


===Procedures===
===Procedures===
*Label the Covaris microtubes.  
*Label the Covaris microtubes. Open the caps and leave tubes on 96-well rack.
*Open the cap and UV-treat the tubes and pipettes for 15min.
*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)
   ---------------------------------------------------------------------------------------------------
   ---------------------------------------------------------------------------------------------------
   1-S         sam080610_2-1_2h    0.685        100          3.43              0      68.5ng
   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

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).