Hello all,
Im trying to Clone a 6KB gene from flower cDNA.Any suggestions before i actually start the work?Im not sure if its gonna be easy amplyfing a 6kn fragment.Is there a good solution to clone this?
Thanks!
0
3 replies to this topic
#2
mdfenko
-
- Active Members
-
- 2,247 posts
an elder
78
Excellent
Excellent
Posted 07 March 2013 - 05:49 AM
use a high fidelity proofreading polymerase in your pcr
Edited by mdfenko, 07 March 2013 - 05:49 AM.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#3
praveen921
-
- Active Members
-
- 7 posts
member
0
Neutral
Neutral
Posted 08 March 2013 - 06:56 AM
Hi i used two polymerases in separate reactions to amplify the 6KB gene
1.Pfu Ultra II DNA Polymerase
2.PrimeSTAR HS DNA Polymerase
I followed the PCR conditions they mentioned in the protocol,but i wasnt able to amplify the gene.What should i do now?
thanks!
1.Pfu Ultra II DNA Polymerase
2.PrimeSTAR HS DNA Polymerase
I followed the PCR conditions they mentioned in the protocol,but i wasnt able to amplify the gene.What should i do now?
thanks!
#4
mdfenko
-
- Active Members
-
- 2,247 posts
an elder
78
Excellent
Excellent
Posted 08 March 2013 - 12:20 PM
check that your primers are appropriate.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
