Talk:Primer (molecular biology)
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[edit] Melting temperature versus annealing
I'm not familiar with this process, but from what I know of annealing, I'm not sure that "melting temperature" is the correct equivalent. --Michael Snow 21:48, 12 July 2005 (UTC)
It's not really about one temperature, but a shifting equilibrium: from annealed to separated and back. So when you lower the temperature, the primer anneals. When you heat it up again it 'melts' (separetes from the dna), but not all at once.
I am a molecular biologist. I modified the optimal length (20bp) and melting temperature (60C) according to my daily experience. I have very good experience with primer3 in picking primers working in experiments later. --Untergasser 14:33, 4 July 2006 (UTC)
- I think Melting temperature and annealing are similar but not exactly the same thing. Melting temperature refers to breaking apart the hydrogen bonds between two DNA/RNA strands. This depends on the length and makeup of each individual strand, as G-C bonding involves a with triple hydrogen bond, and A-T bonding involves a double hydrogen bonds. So Melting temperature is a temperature at which 50% of multiple copies of the same species of DNA/RNA are breaking apart into single stranded molecules. Annealing is the temperature at which short primers hybridize or bind to their complementary sequence in another DNA/RNA molecule which is not necessarily the same length. This will be at a temperature less than the Melting temperature.
- See Polymerase chain reaction for more information --Zven 02:58, 29 August 2006 (UTC)
[edit] Merge proposal
The Polymerase chain reaction article is too big! I propose moving the primer design information here, if it isn't already redundant. I also propose creating a new page PCR optimization and moving all the PCR optimization information there as well. -Madeleine 00:44, 23 March 2007 (UTC)
