Team:Calgary/21 July 2010

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Wednesday July 21, 2010

Raida:

Objective: PCR out native MalE/ MalE 31 protein out of E.coli Purpose: Testing Mal E/ Mal E31 primers to get 1188 bp long gene of interest. Primers that are used are gene specific and anneals right on the start of the Mal E gene.

Tube Labels containing 1.0 μL of 10 mM DNA

  • 1- p1H - C1
  • 2- p1H - C1
  • 3- p1H - C2
  • 4- p1H - C2
  • 5- p31H - C1
  • 6- p31H - C2
  • 7- p1ΔSS - C1
  • 8- Master Mix

PCR Conditions

  • Cycle 1: 94°C 2:00 min
  • Cycle 2 * 36 cycles:
    • Stage 1:94°C for 1:00 min
    • Stage 2:52°C for 0:45 min
    • Stage 3:72°C for 1:15 min
  • Cycle 3: 72°C for 10:00 min
    • Holding Temp: 4°C


Himika

Today I miniprepped the I0500-B0034 construct from yesterday. One of the constructs gave good concentration and good values for protein and RNA contamination. I also restriction disgested the constructs using EcoRI and PstI. These restriction digests were also ran on 1% agarose gel for 45 minutes at 85 volts. The bands on the gel match our expectation. The insert that was cut using the restriction enzymes was about 1200 bp which means that the I0500 got inserted into the plasmid.

No notebook page exists for this date. Sorry!