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