Team:Warsaw/Calendar-Main/11 July 2008

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(Difference between revisions)
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<p>'''Preparation of construct A-omega'''<br>
<p>'''Preparation of construct A-omega'''<br>
-
1. PCR <br>
+
1. PCR A<br>
 +
template DNA - pKS-A4 1ul
 +
primer APNot - 2 ul
 +
primer ALhomo2 - 2 ul
 +
Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 ul
 +
dNTPs - 1 ul
 +
Pfu turbo - 0.5 ul
 +
H2o - 38.5 ul
 +
 
 +
program:
 +
1. 95oC 3'
 +
2. 95oC 30"
 +
3.62oC 45"
 +
4.72oC 45"
 +
5.72oC 10'
 +
6. keeping in 4oC
</p>
</p>

Revision as of 15:48, 26 September 2008

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Preparation of constructs with OmpA protein fusions
1. Colony PCR on colonies from plates with transformations OmpA_alpha.
2. Confirmed transformant colonies inoculated to liquid LB with kanamycin.
Cloning of protein Z DNA to OmpA constructs
1. 2 colonies was inoculated to liquid LB broth with kanamycin

Preparation of construct A-omega
1. PCR A
template DNA - pKS-A4 1ul primer APNot - 2 ul primer ALhomo2 - 2 ul Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 ul dNTPs - 1 ul Pfu turbo - 0.5 ul H2o - 38.5 ul program: 1. 95oC 3' 2. 95oC 30" 3.62oC 45" 4.72oC 45" 5.72oC 10' 6. keeping in 4oC