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Why do DNA primers have fluorescent markers in PCR

..Can someone please explain
Reply 1
Avatar for Mockery
Mockery
17
It's quite simple really, after anelling of the primer to the target DNA, transcription occurs to create the new strand of DNA and because the primer that was used had a fluorescent tag, the DNA strand itself retains the tag.

This allows for DNA profiling and identification of the important loci via electrophoresis methods :smile:
Reply 2
Avatar for Eloades11
Eloades11
17
Original post by mohamedsuleman94
..Can someone please explain


They're able to detect the specific target sequence, meaning non specific products aren't quantified. It also allows multiplex reactions to occur, this is basically identifying any duplications or deletions in a large gene. Beaten to it though :tongue:
Original post by Eloades11
They're able to detect the specific target sequence, meaning non specific products aren't quantified. It also allows multiplex reactions to occur, this is basically identifying any duplications or deletions in a large gene. Beaten to it though :tongue:


Original post by Mockery
It's quite simple really, after anelling of the primer to the target DNA, transcription occurs to create the new strand of DNA and because the primer that was used had a fluorescent tag, the DNA strand itself retains the tag.

This allows for DNA profiling and identification of the important loci via electrophoresis methods :smile:


Thanks and I I think I've got it but (elodes 11) by they're do you mean dna polymerase can detect the target strand to be replicated?
Reply 4
Avatar for Eloades11
Eloades11
17
Original post by mohamedsuleman94
Thanks and I I think I've got it but (elodes 11) by they're do you mean dna polymerase can detect the target strand to be replicated?


Not quite, if the primer sequence isn't complementary to the strand then it won't bind (unless every other base is and the primer is large, which induces a point mutation). The primers recognise and bind to the existing template strand, the DNA polymerase simply recognises the dsDNA with the primer, and replicates the rest of the strand. Does this help? :smile:
Original post by Eloades11
Not quite, if the primer sequence isn't complementary to the strand then it won't bind (unless every other base is and the primer is large, which induces a point mutation). The primers recognise and bind to the existing template strand, the DNA polymerase simply recognises the dsDNA with the primer, and replicates the rest of the strand. Does this help? :smile:


Yes, thanks a lot!

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