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-   -   Polymerase chain reactions? (http://www.molecularstation.com/forum/pcr-polymerase-chain-reaction-forum/84124-polymerase-chain-reactions.html)

Crystal 11-24-2011 09:16 PM

Polymerase chain reactions?
 
Primer 1: 5'-ATGAAGATCTTCTTATTCTTAGCT-3'
Primer 147: 5'-CTAGTAATAGCCAAAAGTCAC-3'

Primer 1 primes synthesis of the 5' to 3' strand of an oat cdna/gene and encodes for the amino acids appearing at the N-terminal end of the corresponding polypeptide. Primer 147 primes synthesis of the 3' to 5' strand of an oat cdna/gene and encodes for the amino acids appearing at the C-terminal end of the polypeptide. They polypeptide encoded by this gene is 147 aas in length.

A) The PCRs using Plasmids with primer 1 and 147 were "positive controls". What is the expected size of the DNA fragments amplified in PCR using these primers and plasmids? Why?

B) Assume that a PCR using the combination of primer 1 and 147 plus oat genomic DNA resulted in the applification of a DNA fragment of 450bp larger than the "positive controls" above. With respect to the structure to this relevant gene, what does this result indicate? Explain?

davidflora 12-20-2012 07:59 AM

Re: Polymerase chain reactions?
 
[Only registered and activated users can see links. Click Here To Register...]can add a nucleotide only onto a preexisting 3'-OH group, it needs a primer to which it can add the first nucleotide. This requirement makes it possible to delineate a specific region of template sequence that the researcher wants to amplify. At the end of the PCR reaction, the specific sequence will be accumulated in billions of copies (amplicons).


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