IGEM:Cambridge/2008/Notebook/Voltage/2008/07/25: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Autocreate 2008/07/25 Entry for IGEM:Cambridge/2008/Notebook/Voltage) |
|||
| Line 5: | Line 5: | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
== | ==Double Terminator== | ||
The DNA was extracted from lanes 3&4 of the gel and extracted using the Zyppy Gel Extraction kit. | |||
The extracted DNA was then PCRed using 34 cycles, and then 10μL was froxen at -20°C. | |||
8μL of the PCR product was cut with both EcoR1 and Pst1, as was 5μL of the death plasmid PSB4C5, using the fast digest protocol. | |||
The gene was then ligated into the plasmid backbone, using the fast ligation kit, from Finnzymes. | |||
500μL of normal TOP10 cells were made competent, and then transformed with the 4μL of the new plasmid, and grown on chloramphenicol overnight. 1μL of Puc19 was used as a control. | |||
==K+ assay for E.coli== | |||
Revision as of 12:58, 29 July 2008
 ........... Electric Output
|
<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> |
Double TerminatorThe DNA was extracted from lanes 3&4 of the gel and extracted using the Zyppy Gel Extraction kit. The extracted DNA was then PCRed using 34 cycles, and then 10μL was froxen at -20°C. 8μL of the PCR product was cut with both EcoR1 and Pst1, as was 5μL of the death plasmid PSB4C5, using the fast digest protocol. The gene was then ligated into the plasmid backbone, using the fast ligation kit, from Finnzymes. 500μL of normal TOP10 cells were made competent, and then transformed with the 4μL of the new plasmid, and grown on chloramphenicol overnight. 1μL of Puc19 was used as a control. K+ assay for E.coli | |
