Team:Paris Bettencourt/Notebook/TB-ception/Tuesday 10th September.html

TB-ception

ASDF

Tuesday 10^th September

• PCR for the linearized backbone: template - pSB1C3 (1 µL), primers (SB-prep-2Eb, SB-prep-3P-1; 2.5 µL), HE buffer (10 µL), dNTPs (1 µL), polymerase (0.5 µL), dH2O (32.5 µL).

program: 94.0 (30 s) - 94.0 (10 s), 55.0 (1 min), 72.0 (2 min), 72.0 (3 min) - 30 cycles.
• touchdown PCR for the new backbones:

Template	Primers
pACYC	pDCA2 and pDCA
pET	pDCo4-F and pDCo4-R
pCDF	pDCA1 and pDCA
pColA	pDCA3 and pDCA

program: 95.0 (10 s) - 95.0 (30 s), 60.0 (30 s), 59.0 (30 s),58.0 (30 s), 56.0 (30 s), 55.0 (30 s), 72.0 (30 s) - 72.0 (10 min)- 30 cycles.