Revision as of 07:27, 23 September 2016

Friday 2th ^th August

By Mahnaz

Q5 PCR and phusion PCR were performed on plasmids with the following protocol:

For each 50μl of reaction, mix the following reagents:

Mix gently and aliquot 50 μl of the mix into the PCR tubes on ice. Perform PCR as follow:

Primers used were:

Matrix	gblock 1.1 in pUC19	gblock 1.2 in pJET	gblock 2.1 in pUC19	gblock 2.2 in pUC19
Primers	iPS140 and iPS120	iPS121 and iPS122	iPS123 and iPS124	iPS125 and iPS84
Tm	72°C	72°C	72°C	72°C
t	30 sec	30 sec	30 sec	30 sec

By Mahnaz

PCR products obtained were cleaned up by using the NucleoSpin Gel and PCR Clean-up kit protocol.

By Mahnaz

Sample PCR reaction	Q5 reaction Concentration (ng/µL)	phusion reaction Concentration (ng/µL)
gblock 1.1	161.88	263.81
gblock 1.2	218.01	209.24
gblock 2.1	101.16	185.47
gblock 2.2	113.88	138.87

@@ Line 87: / Line 87: @@
 {| class="wikitable"
-!Sample phusion reaction
+!Sample PCR reaction
-!Sample Q5 reaction
+! Q5 reaction Concentration (ng/µL)
-!Concentration (ng/µL)
+! phusion reaction Concentration (ng/µL)
 |-
 |gblock 1.1 <div id=" gblock 1.1 "></div>