PCR help

[wolfmiffy]

hi there,

If I am going to do a PCR amplification of a region in the E. coli genome. I wonder if the first PCR melting temperature should be the same as 95 degree or should it be changed to a much higher temperature? If anyone knows any protocal of doing PCR on genomic DNA plz let me know the links. Thank you!

[Aga]

95 should be fine. What do you mean by genomic DNA PCR? Is it just classic PCR where the template is the genomic DNA and amplified DNA fragment is just several hundred of bp long?

[wolfmiffy]

yuppppppppppppppper thanks

[ramnivas]

Ya there is not such specificity its just simple PCR programme. You can decide extenstion time according to the length of amplicon 1 min/1kb

[elpachito]

cool, we are learning about pcr in microbiology.