Hi Govardhan,
As also mentioned by Thanes .....the amplification of a PCR product using either forward/reverse primer is possible.....since there will be a template and a primer the Polymerase will work anyway....but you will get extremely low amount of product in each cycle....'that means it will not be a regular Polymerase 'chain reaction'....or you will not get double the number of molecules in every cycle....As a consequence you will not get the same amount of product say in 35 cycles compared to when you use both the primers......BUT YOU CAN DO PCR EVEN USING ONE OF THE PRIMER....
There are some approaches that uses a single primer and PCR for different applications...PCR based Cycle sequencing is one of them.....
Hi Govardhan...
Single primer when used in PCR can give amplified products...provided it has binding sites in the template, that are present in the inverse orientation and within a amplifiable distance defined by the extension time. Many single primer based techniques are in routine use..such as RAPD, ISSR etc..
Thank you sir
regarding this any information please send me sir
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Hi Govardha
I'll definitely send you some good articles ..on these techniques
Hi,
What do u mean by single primer? Is it Single set of primer which includes forward and reverse? or only of either forward or reverse primer? In the first case, you will get a complete band and can be observed in gel. In the second case, the amplification still happens but only gives one strand depends on the primer used and hence the band cannot be seen in the gel due to the fact that the intercalating dye (eg. EtBr) binds in between the double strands.
thank you
i mean either forward or reverse primer
is there any application for this type PCR
you will not get a pcr product if you use only forward or only reverse primer. The amplification will result only if both forward and reverse primers are used
Hi Govardhan,
As also mentioned by Thanes .....the amplification of a PCR product using either forward/reverse primer is possible.....since there will be a template and a primer the Polymerase will work anyway....but you will get extremely low amount of product in each cycle....'that means it will not be a regular Polymerase 'chain reaction'....or you will not get double the number of molecules in every cycle....As a consequence you will not get the same amount of product say in 35 cycles compared to when you use both the primers......BUT YOU CAN DO PCR EVEN USING ONE OF THE PRIMER....
There are some approaches that uses a single primer and PCR for different applications...PCR based Cycle sequencing is one of them.....
Just to complement... Using a set of specific primer, the theoretical equation would be something like that
Qtt = A0 * 2^(nc) (Qtt = quantity of DNA in copies, A0 = initial amount of dna in copies, nc = number of cycles)
And with only one primer (either forward or reverse), you'd have:
Qtt = A0 * (nc+1)
That means that, with an initial amount of 1000 copies, at the end of 35th cycle you would have 34359738368000 copies with a set of primers (that is, if your reaction goes on 100% efficiency until the end), while with only one primer you'd have only 36000 copies.
Besides, I believe that using only 1 primer would not give you an amplification product with a specific size (because there is not a second one to limit the size, so DNA polymerase just keeps extending that one strand until you change the temperature).
Hi Guys
single primer PCR is a technique that you can use one specific primer (R or F) and specific conditions and reactions to get single strand product and it is useful when you need to identify the transposon region after mutation and you will set PCR in 2 or 3 round at the same time and the final product you get will be a template to sequence it and get an exact region of your transposant. regards
HI
If u use single primer i think it wont give u any amplification product if ur primer is 18 - 20 oligomer. If u use short sequence primer such as 8-10 oligomers which is in the case of RAPD ur single primer will act as both forward and reverse and u will get multiple band which will be use to study the polymorphism, whereas if use single primes for a particular gene or target u wont get the specific product.
Single primer polymerase chain reaction currently well used to amplify unknown DNA fragments adjacent to a short stretch of known sequence for DNA sequencing especially ITRs.
Hi,
I just stumbled over this and found this old paper describing what you want to do:
Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus
http://www.pnas.org/content/85/20/7652.full.pdf
Basically they use 2 primers, but 1 in drastic excess (1:100-500) so that first they get an amplification of their desired amplicon and then when the low concentration primer is "used up" mainly ssDNA is produced in the reaction.
So whoever stumbles on this next might find that technique helpful.
maybe this can help
and yes, you will get the pcr product even with single primer
Many technique use single primer sequence. This primer use in PCR reaction as forward and reverse primer if it get binding site such as RAPD, ISSR, SCoT and other
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