The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
 iGEM 2011
|
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>
|
Objectives
- See resctriction in gel to perform ligation.
- First PCR with standard primers for Isothermal assembly.
Gel
- Lane 1 Ladder O'Gene Ruler mix
- Lane 2 B0015 EcoR1/Pst1
- Lane 3 pSB1T3
It is a little bit dirty but usefull for Ligation.
Ligation performed by Vladimir
PCR with Standard primers for isothermal assembly
Using Cambridge protocol for primers and backbone assembly.
| Reagents
|
Insert of P1A7
|
Insert of P1A7
|
Backbone pSB17 P1A7
|
Backbone pSB17 P1A7
|
pUXDNA Insert Control
|
pUXDNA Backbone Control
|
| Buffer Phusion HF 10x
|
10μL
|
10μL
|
10μL
|
10μL
|
10μL
|
10μL
|
| 8mM dNTPs
|
5μL
|
5μL
|
5μL
|
5μL
|
5μL
|
5μL
|
| Primer Fwd
|
5μL
|
5μL
|
5μL
|
5μL
|
5μL
|
5μL
|
| Primer Rev
|
5μL
|
5μL
|
5μL
|
5μL
|
5μL
|
5μL
|
| DNA
|
1μL
|
1μL
|
1μL
|
1μL
|
1μL
|
1μL
|
| Phusion
|
.5μL
|
.5μL
|
.5μL
|
.5μL
|
.5μL
|
.5μL
|
| H2O
|
23.5μL
|
23.5μL
|
23.5μL
|
23.5μL
|
23.5μL
|
23.5μL
|
| Total
|
50μL
|
50μL
|
50μL
|
50μL
|
50μL
|
50μL
|
|
iGEM 2011
