Biochemistry 2280A Lecture Notes - Lecture 22: Agarose Gel Electrophoresis, Ethidium Bromide, Agarose
12 Dec 2017
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How do we get enough yfg to be able to clone. Exponential amplification of any dna from a source in which it is found as little as one. You selectively amplify yfg from a source of proteins. Required reagents for pcr: template dna, two oligonucleotide primers which flank yfg, dntps, dna polymerase. Hybridize primes at 50, dna synthesis of primers with dna polymerase dntps, at 72 degrees make dna copy. Fi(cid:396)st (cid:272)y(cid:272)le p(cid:396)odu(cid:272)es (cid:1006), se(cid:272)o(cid:374)d (cid:1008), thi(cid:396)d 8 . Number copies after n cycles is 2^n with 1 template molecule. Thermocyclers that oscillate between the three required temperatures. Gel electrophoresis separates dna on the base of size. Dna is sieved through a matrix that contains small pores being pulled by an electric field. After melting it hardens into a form like jell-o. It has small pores through which the dna can pass through. The agarose gel is in a chamber submerged in buffer, an electric field is then applied.
