Sam:LabNotes/Microbiome-new/2010-2-4

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Covaris shearing for mock E.coli gDNA sample with carrier - at Harvard[edit]

Backgournd[edit]

  • Using sheared E.coli DNA in verious amount to test the low-limit of DNA requirement for Solexa libarary construction.
  • tRNA carrier was used to avoid the loss the DNA on tubes.

Precedures[edit]

  • E.coli DNA dilution
    • 1/10 dilution => 56.2ng/uL
    • 1/100 dilution => 7.8ng/uL, 7.8ng/uL -> 1/7.8 => 1 ng/uL
  • tRNA concentration: 121ng/uL
        200ng                 50ng           10ng                2ng                0ng
----------------------------------------------------------------------------------------
DNA  200/56.2 = 3.56      50/7.8 = 6.4     10/7.8 = 1.28      2/1 = 2.0              0
tRNA 200/121  = 1.65     350/121 = 2.9    390/121 = 3.22   398/121= 3.29    400/121= 3.3
H2O            44.79              41.7             45.5            44.71            46.7
----------------------------------------------------------------------------------------
               50.00              50.0             50.0            50.0             50.0

Prepare four replicates for each type of samples in 0.5mL non-sticky tubes(Ambion)
  • Ship to Sara (Havard) at RT by FedEx standard overnight
    • Tracking no.: 4027-6591-9356
  • Covaris shearing condition
duty cycle: 10%
intensity: 5
bursts per cycle: 200
180 seconds