Biochemistry 2280A Lecture Notes - Lecture 23: Genomic Library, Sequence Alignment, Plasmid

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Shotgun genome sequencing (used to sequence small genomes) Steps: creating a genomic dna library, fragment dna and determine the sequence of each fragment, aligning the independent sequences into a continuous sequence. A collection of cloned dna fragments that represents all of the dna in an. Genomic dna library (all genes in the genome) organism"s genomes. All genes are equally present in dna library, but not equally present in cdna library: no introns because some are transcribed more than others in to rna. Sequencing the genome of h. influenzae: the first genome sequenced. Dna was sonicated (fragmenting using high frequency sound) in to fragments with blunt. Fragments were electrophoresed, those in size range of 20kb (best) were purified from gel. Fragments were ligated into plasmids and bacteria were used to make clonal library. They chose 20 000 sequences from the library at random and sequenced. Step 2: sequence experiments the ends of each one.

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