IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 RNA Decoder/2010/06/21

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June 21, 2010

Today we started to clone out our genes that are needed for the decoder. We did cloning for MicA, MicF, Hfq, OmpA, and OmpF. The PCR procedure we followed was a 40 uL reaction:

  • 10X Buffer 4 (uL)
  • 10mM dNTPs 1 (uL)
  • Template 2.5 (uL)
  • Forward primer .4 (uL)
  • Reverse primer .4 (uL)
  • Polymerase .4 (uL)
  • dH2O 31.3(uL)
  • Total 40 (uL)

The template used was the MG1655 E.Coli genome.

We then ran a electrophoresis gel for the reactions and saw bands that were the desired sizes for Hfq, OmpA, and OmpF. MicA and MicF had no band in their lanes.

We took concentration readings for the three genes that were acquired from the PCR:

  • OmpA 34.29 ng/uL
  • OmpF 76.75 ng/uL
  • Hfq 36.57 ng/uL