Cloning at 6 kb gene - (Mar/07/2013 )
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?
use a high fidelity proofreading polymerase in your pcr
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?
check that your primers are appropriate.