BIOL 200 Study Guide - Final Guide: Palindromic Sequence, Proper Length, Telomere
22 Jun 2016
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Dna-pk and ku80/ku70 bind to broken ends of dna. Ends digested by exonuclease, 3" single stranded ends overhang. Reca or rad51 mediated strand invasion (crossing over) forms watson crick base pairs. 3" end of invading strand is extended by dna pol until the both displaced single strands base pair with each other. Polylinker region, di erent sites for di erent restriction enzymes endonuclease makes staggered cuts (gaa ttc for ecor1) palindromic sequence. Cuts between g and a forms sticky ends mix the plasmid with e. coli, and introduce ampicillin. Taq dna polymerase (heat resistant and stable) primers (oligonucleotides or oligos) short dna molecule dntps. Steps: 95 degrees c - denature dna, 60 degrees c - anneal primers, 72 degrees c - dna extension, repeat 20-40 times. After 2-3 cycles you get the proper length. Dna template dntps (100mm) ddntps (1mm) (chain terminator) removes the 3" oh. Do 4 reactions but use a di erent ddntp each time (a, c, g, t)
