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 - 95oC for 30 secs,
30 cycles - 95oC for 30 sec, 60oC for 45 seconds,72oC for 1 min
The results show that 6L has failed whilst the XylR has sucessfully been ligated.