VPB 321: Animal Biotechnology (2+1)

BAC TO BAC SEQUENCING

• These pieces are fingerprinted to give each piece a unique identification tag that determines the order of the fragments.
• Fingerprinting involves cutting each BAC fragment with a single enzyme and finding common sequence landmarks in overlapping fragments that determine the location of each BAC along the chromosome.
• Then overlapping BACs with markers every 100,000 bp form a map of each chromosome.
• Each BAC is then broken randomly into 1,500 bp pieces and placed in another artificial piece of DNA called M13. This collection is known as an M13 library.
• All the M13 libraries are sequenced. 500 bp from one end of the fragment are sequenced generating millions of sequences.
• These sequences are fed into a computer program called PHRAP that looks for common sequences that join two fragments together.