测序及建库专题.pptVIP

Sequencing Two ways for sequencing: 1. DNA molecules (radioactively labeled at 5’ termini) are subjected to 4 regiments to be broken preferentially at Gs, Cs, Ts, As, separately. (Maxam and Gilbert chemical method, not widely used) 2. Chain-termination method (Sanger’s method, widely used) Chain-termination method ddNTPs are chain-terminating nucleotides: the synthesis of a DNA strand stops when a ddNTP is added to the 3’ end Shotgun sequencing of a bacterial genome The shotgun strategy permits a partial assembly of large genome sequence Assembly Step 1: form contigs (A single contig is abou