User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/10/17: Difference between revisions
From OpenWetWare
Line 21: | Line 21: | ||
The backbone vector has BSMBI cut site within the hygromycine resistance gene. I was removed though [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/10/10 Site Directed Mutagenesis] | The backbone vector has BSMBI cut site within the hygromycine resistance gene. I was removed though [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/10/10 Site Directed Mutagenesis] | ||
---- | |||
'''PCR Reactions''' | |||
{| | |||
| align="center" style="background:#f0f0f0;"|'''Part''' | |||
| align="center" style="background:#f0f0f0;"|'''BD002''' | |||
| align="center" style="background:#f0f0f0;"|'''HPk''' | |||
| align="center" style="background:#f0f0f0;"|'''CMV''' | |||
|- | |||
| DNA Template (µL)||0.1||0.1||0.1 | |||
|- | |||
| Forward Promer (µL)||1||1||1 | |||
|- | |||
| Reverse Primer (µL)||1||1||1 | |||
|- | |||
| 2X GoTag (µL)||50||50||50 | |||
|- | |||
| dH2O (µL)||47.9||47.9||47.9 | |||
|- | |||
| Tolal (µL)||100||100||100 | |||
|- | |||
| | |||
|} |
Revision as of 15:09, 24 October 2013
PcTF Subcloning in E. coli | <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> | |||||||||||||||||||||||||||||
Golden Gate Assembly of BD002 to Replace HsvtkTATA with CMV and HPK PromotersDesired Constructs: BD003: 5XGAL4-Spacer-CMV-Kozak-AMCyan-AmCyan-NLS-Stop BD004: 5XGAL4-Spacer-HPK-Kozak-AMCyan-AmCyan-NLS-Stop Existing construct is: 5XGAL4-Spacer-HsvTK-Kozak-AMCyan-AmCyan-NLS-Stop in V0200 mammalian vector Amplification of CMV and HPK from KAH187 and KAH184 respectively. The backbone vector has BSMBI cut site within the hygromycine resistance gene. I was removed though Site Directed Mutagenesis PCR Reactions
|