Team:Stockholm/10 August 2010

From 2010.igem.org

(Difference between revisions)
(New page: {{Stockholm/Top2}} == Mimmi == === yCCS and SOD === ==== redoing the Site-Directed Mutagenesis ==== {| ! Mix | (µl) | X2 X2 | rowspan="5" width="100" | ! '''yCCS A''' | rowspan=...)
(→Mimmi)
Line 195: Line 195:
| 10
| 10
|}
|}
 +
 +
==Andreas==

Revision as of 12:44, 11 August 2010

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

↳   Notebook Safety Protocols



Contents

Mimmi

yCCS and SOD

redoing the Site-Directed Mutagenesis

Mix (µl) X2 X2 yCCS A SOD A conditions
sH2O 40 80 yCCS_mut_F SOD_mut_F time °C
dNTPs 1 2 yCCS_mut_R SOD_mut_R 30s 95
F primer 1 2 pSB1C3 pSB1C3 30s 95 )
R primer 1 2 ~3.5kb ~3.2kb 30s 55 > 22 cycles
DNA 1 2X1 4m 68 )
Pfu buffer 5 10 oo 10
Pfu turbo 1 2
tot 50
SOD Primers
SOD_mut_F 262.55µl H2O --> 100µM
SOD_mut_R 372.64µl H2O --> 100µM
SOD_mut_F 3µl + 24.15µl H2O --> 125ng/µl
SOD_mut_R 3µl + 24.42µl H2O --> 125ng/µl


MITF

Amplifying

Mix phusion (µl) /4 Mix Pfu turbo (µl) /4 Primers
sH2O 67 sH2O 77 MITF_F
F primer 5 F primer 5 MITF_R
R primer 5 R primer 5
dNTP 2 dNTP 2 conditions
5X buffer 20 10X buffer 10 time °C
Phusion pol. 1 Pfu pol. 1 2m 98
tot 100 25 tot 100 25 30s 98 )
30s 40-55 > 5 cycles
1m30s 72 )
30s 98 )
30s 65 > 25 cycles
1m30s 72 )
10m 72
oo 10

Andreas

Retrieved from "http://2010.igem.org/Team:Stockholm/10_August_2010"