User:Tara K. Luckau/Notebook/Team ConGen/2010/11/04

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Contents

Cycler Test with new silicone mats

Set-Up

  • received sample of silicone mats (got 5! yay for free stuff!) from BioExpress, Catalog# T-3163-2
  • 10µL in each well of full-skirt PCR plate, PCR seal, silicone mat
  • Data: 20101104_TKL_CyclerTestBEMat.tad

Results

  • evaporated: A4, A5, A6, H6, A7, A8, H9
  • will probably use, anyway ...

PCR Scun2 F-R

  • optimization PCR with 3 variables: DNA concentration, Buffer, Temperature
  • determine lowest DNA concentration (1, 1/2, 1/10, 1/100) that works on PCR ... determine NanoDrop concentration ... use that value as goal concentration for all future PCR

  • DNA: from mass extraction (that Tina did), SCOC SYR9 X04
  • Ran out of Big MM for Buffer G, so H12 only has about 4µL
  • used new BioExpress silicone mats (catalog# T-3163-2)

NanoDrop SCOC SYR9 X04

  • this is the DNA sample used in the optimization gradient PCR, 4 November 2010
Sample ng/µL A260 A280 260/280 260/230
Zero 0.73 0.015 0.023 0.62 1.67
1x -4.43 -0.089 -0.050 1.79 0.78
1/2x -0.93 -0.019 0.003 -6.68 -0.01
1/10x 17.07 0.341 0.190 1.80 0.04
1/100x 43.84 0.877 0.548 1.60 0.07
  • Only 1x and 1/2x gave positive concentration, but they are consistent with each other (2 x 1/2x ≈ 1x)



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