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