1 October 2008

Wet Lab

PCR

• Today we carried out a Single colony PCR to check a the ligation of parts into biobrick vectors. The image below summaries the results of the PCR. The labels correspond to the following:
  • 6L - LipA-Elastin into AK3 vector
  • X- XylR into AK3 vector
  • Pos-Positive control of AmyE 5' (577bp)
  • Neg- Negative control, no DNA template.

• The following reaction conditions were used with the Pfu ultra enzyme:
  • 1 cycle - 95^oC for 30 secs,
  • 30 cycles - 95^oC for 30 sec, 60^oC for 45 seconds,72^oC for 1 min

• The results show that 6L has failed whilst the XylR has sucessfully been ligated.