how to design pcr primers - (Jul/07/2008 )
hi
i want to design pcr primers for my project to amplify desired gene sequence
fast pcr pcr designing software is available with me
please tell me how to design pcr primers with fast pcr softwre
how to take care with troubles like primer dimer formation ,and all
please tell me what are all precautions and troubleshooting for primer designing for required dna sequence to be amplify
thanks in advance
you cannot take all possible precautions or troubleshoot before encountering problems with PCR...
why don't you take a look at the Manual of your program first, or use a simpler web-based form like Oligoperfect. If it works, it works, if it doesn't, troubleshoot, what can I say...
note that at the time I designed my first primers, I did it with oligoperfect, and it just worked fine...
why don't you take a look at the Manual of your program first, or use a simpler web-based form like Oligoperfect. If it works, it works, if it doesn't, troubleshoot, what can I say...
note that at the time I designed my first primers, I did it with oligoperfect, and it just worked fine...
thank you .,i will try this
why don't you take a look at the Manual of your program first, or use a simpler web-based form like Oligoperfect. If it works, it works, if it doesn't, troubleshoot, what can I say...
note that at the time I designed my first primers, I did it with oligoperfect, and it just worked fine...
please tell me briefly how to proceed for designing primers (from sequnce to primers)
it will be very helpful for me
i need help
thank you
First check in the literature if somebody do a similar work and use the primers given for start. But if has to do it from scratch. Check in pubmed or similar website the secuence of the gene. Choose the region of interest. Go to your software or use Primer3 (good tool and is free) Paste the secuence and enter the size of the amplicon, size of the oligo, etc. The program will give you several pairs of primers. Take the first best that fit and go to Blast nucleotide (pubmed) and enter the secuence of each primer. This will tell you how specific is the primer secuence. All the process will take you several hours, but you will be sure that what you design will be functional.
thank you
thank you
your welcome...we are here to help as much as we can...