Team:Warsaw/Calendar-Main/2 June 2008

From 2008.igem.org


	Attribution analyzer / Interactive list of topics

Rifampicin test #3

Piotr

Strain	Inducer	Number of colonies on rifampicin	OD
TOP10	none	9	2.66
TOP10 pMPM-T5-AID	none	15	2.5
TOP10 pMPM-T5-AID	0.1% Arabinose	42	2.27
TOP10 pTrc99a-AID	none	12	2.41
TOP10 pTrc99a-AID	0.5 mM IPTG	35	1.98

Conclusion: AID seems to work, induction of AID may reduces the growth rate

Preparation of pMPMT5-AID+AIDT7 construct

Michał K.

PCR for AID transcriptional fusion with translational fusion AID+T7 RNA-polymerase.
Primers: AIDlNrH and T7pXbSal
Template DNA: translational fusion AID+T7 RNA-polymerase (pMPMT5+AID-T7)
Elongation temperature: 55°C
Time: 4 minutes
Number of cycles: 20 and 25

Blue/white and rifampicin test #1

Michał L., Ewa, Marcin

Transformation of E. coli TOP10 with pZC320.
Plating transformants on LB + ampicillin (30 μg/mL) + X-gal + IPTG.