CMMB 461 Lecture Notes - Lecture 4: Restriction Digest, Complementary Dna, Chromosome
16 Sep 2018
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Human chromosomes (23 pairs) range in size from 10 100"s million bases long. Note that sequencing reads are still up to 800 base pairs using the abi 3700 dna analyzer. Sequencing reads is another way of saying sequencing reaction. Strategy requires breaking the genomes into smaller fragments or clones and sequencing these fragments this is the shotgun approach. In this approach, pellets are fired at dna which hit the dna at random spots, causing the genome to be fragmented into different sizes. Purpose is to store and make many copies of a dna fragment of interest in e. coli. Required to initiate transcription, and further, translation. Used to cut the vector and insert gene. It is essentially a short segment of dna which contains up to 20 restriction sites1. This site is also known as a polylinker. Helps purify protein and tag the gene.
