Team:Stockholm/17 September 2010

From 2010.igem.org

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==Johan==
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PCR again, to see if the 900 nt-band or the 700 nt-band is correct
 +
 +
0,5 µl pol
 +
 +
0,5 µl dNTP
 +
 +
5 µl 5x buffer
 +
 +
1,5 µl for primer
 +
 +
1,5 µl rev primer
 +
 +
16 µl H2O
{{Stockholm/Footer}}
{{Stockholm/Footer}}

Latest revision as of 01:37, 28 October 2010

Home Project Idea Lab Work Results Modelling Team       Follow us on and        2010.igem.org

↳   Notebook Safety Protocols


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

Johan

PCR again, to see if the 900 nt-band or the 700 nt-band is correct

0,5 µl pol

0,5 µl dNTP

5 µl 5x buffer

1,5 µl for primer

1,5 µl rev primer

16 µl H2O





The Faculty of Science at Stockholm University Swedish Vitiligo association (Svenska Vitiligoförbundet) Geneious Fermentas/ Sigma-Aldrich/

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