5 replies to this topic

[novagen]

Hi all,
I have designed the primers using the UTR region of the gene. Which template should be used for PCR amplification.Is it genomic DNA or CDNA.

waiting for your reply
Thanx in advance.

[Heir Stylist]

novagen, on Jan 26 2009, 09:12 PM, said:

Hi all,
I have designed the primers using the UTR region of the gene. Which template should be used for PCR amplification.Is it genomic DNA or CDNA.

waiting for your reply
Thanx in advance.

cDNA / genomic DNA should work if the primer binding sites (of the UTR )are close to the ORF. Keep in mind that if you are using Genomic DNA from Eukaryotic organisms, your gene coding regions would be interrupted by introns ( and you have to deal with a larger amplicon compared to the one with the cDNA)


Regards
Avinash
http://aviprem.gq.nu

[lotus]

cDNA should be okay I guess.

[smallcat227]

In my opinion, cDNA, especially in eukaryotic organisms, because of the  introns.

[pcrman]

You did not mention what your purpose is for the PCR? cloning or assessing expression?

[novagen]

hi all,
my purpose is for cloning and then getting it sequenced

Thanks