Biochemistry 2280A Lecture : Polymerase Chain Reaction
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Dna molecules are most easily separated based on difference in length (number of base pairs) Dna is sieved through a matrix of agarose (or acryladmide gel) In gel there are very small pores; the dna goes through these pores. Dna from each pcr reaction is loaded into a well in the agarose gel. The gel is in a chamber in which an electric field can be applied. When an electric current is passed through the chamber, dna fragments are pulled through the agarose matrix and move towards the positive pole. Smaller dna fragments move faster than large dna fragments. Detection of the dna in the gel: stain with ethidium bromide. Expose yor gel to an x-ray film (figre. Introns are a problem in expressing eukaryotic genes in e. coli. Cdna is a replicate dna copy of mrna (c=complimentary) Cdna does not contain introns, and each cdna encodes one gene. Start by making rna form the tissue of interest.