Biochemistry 2280A Lecture Notes - Lecture 21: Reverse Transcriptase, Electric Field, Polynucleotide
30 Dec 2017
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Making cdna: mrna can"t be cloned- must convert it to cdna using reverse transcriptase (enzyme) cdna is a complementary dna copy of the mrna (c=complementary) Treat with alkali to degrade rna- left with cdna (can make cdna double stranded if needed, for our purposes we do not) Fragments of single stranded dna with defined sequence that are made through repetitive chemical reactions: when they are short (20-30 bases) they are referred to as primers or oligos. Polymerase chain reaction (pcr: amplify gene, exponential amplification of any dna from a source in which it is found as little as once (one copy is. Invented by kerry mullis in the early/mid 1980s, became a standard technique 1990 enough for pcr: note: you can selectively amplify yfg from a complex mixture of genes. Required reagents for pcr: template dna (we"re using cdna, two oligonucleotide primers which flank yfg, one hybridizes to 5" side, other hybridizes to 3" side, dntps, dna polymerase.
