Team:Stockholm/17 September 2010

From 2010.igem.org

Revision as of 12:45, 4 October 2010 by Mimmi (Talk | contribs)


Contents

Andreas

Plasmid preps

From 16/9 ON cultures

DNA concentration
Sample Conc [ng/μl] A260/A280
pSB1C3.N-TAT 64.07 n/a
pSB1C3.N-Tra10 46.38 n/a
pSB1C3.N-LMWP 65.77 n/a
pSB1K3.N-TAT⋅SOD⋅His 4 87.83 n/a
pSB1K3.N-TAT⋅SOD⋅His 5 8.464 n/a
pSB1K3.N-Tra10⋅SOD⋅His 5 6.872 n/a
pEX.SOD 31.57 n/a




Mimmi

SOD / yCCS

over expression

  • Start culture
    • 10ml LBAMP + 100µl old culture (8:30)
  • Measure OD=0.6 (11:30)
    • At OD=0.6 add IPTG 1mM
  • Take samples after:
    • 0h
    • 2h
      • take 500µl, spinn down and remove LB
      • resuspend in 50µl SDS buffer
      • Heat in 95°C, 5min
      • Freeze
      • Re-heat in 95°C, 5min
  • Run gel



PhastGel

well sample
Place for picture.jpg
well sample
Place for picture.jpg
1 ladder 1 ladder
2 SOD.his 0h 2 yCCS 1 0h
3 SOD.his 2h 3 yCCS 1 2h
4 his.SOD 0h 4 yCCS 2 0h
5 his.SOD 2h 5 yCCS 2 2h
6 ladder 6 ladder


his.SOD.cTAT

colony PCR

mix (µl) x8 Primers conditions
mastermix 24.5 pSB1_VF2 time °C
DNA 0.5 pSB1_VR 5m 95
tot 25µl 30s 95 )
30s 55 > 30 cycles
1m20s 72 )
10m 72
oo 25