1.3kb PCR only shows primer dimer - (Dec/26/2005 )
hi!
im amplifying a gene of known molecular weight (1.3kb) after designing forward reverse primers.when i run pcr ,im getting d desired band but ofvery weak intensity. besides i can see a very strong band of v low molecular weight in d gel . may be a primer dimer. how do i get the desired band of high intensity as i need to sequence n clone tht fragment . do i need to change the primer concentration or temp of annealing in my pcr? ![]()
-promilasharma-
Try decreasing your annealing temp or increase your Mg-concentration a bit. How big is the low molecular weight strong band? Is it a couple 100 bp or below 100 bp?
How many cycles do you run?
-vairus-
QUOTE (vairus @ Dec 26 2005, 03:31 AM)
Try decreasing your annealing temp or increase your Mg-concentration a bit. How big is the low molecular weight strong band? Is it a couple 100 bp or below 100 bp?
How many cycles do you run?
How many cycles do you run?
im running 30 cycles, n the band is more than 100bp
-promilasharma-