Alkendi Posted February 27, 2007 Posted February 27, 2007 hi all: I am working with universal primers for bacteria and I came a cross some primers that are like follwing: F984GC (position:968 to 984) R1378 (position: 1378 to 1401) this pair of primers is used to amplify 16S rDNA gene. the question is that if the forward primer starts at nucleotide 968 and stops at nucleotide 984 and the reverse primer starts and finishes at different site so, these two segments do not hybridize with each other. since the sense strand segment is different than the antisense strand segment then the amplification using this pair of primer will result in two segments. is what I am saying correct, please tell me if I am wrong or if you can clarify more I would appreciate it. Thanks
CharonY Posted March 1, 2007 Posted March 1, 2007 I am not perfectly sure what you are asking. Using these primers will give you a single product starting from 984 and ending with bp 1401 of the template.
Alkendi Posted March 13, 2007 Author Posted March 13, 2007 May I please ask you if the pair of primer (Forward and Reverse) are work on one templete on the same time. meaning that one goes from 5' to 3' and the other goes from 3' to 5'? thanks for your answer
Bluenoise Posted March 14, 2007 Posted March 14, 2007 No. All DNA is synthesised in the 5' to 3' direction. Meaning that new nucleotides are added onto the 3' end. This is universal in all cases. One primer acts on one strand, the second acts on the complementary strand. that way you gen production of two strands that are complementary to each other.
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