DNA Sequencing

what is the Sanger-Chain Termination Method

uses small amounts of dideoxynucleotides that when incorporated cause termination of primer chain extension

what is shotgun sequencing of genomes

Random DNA fragments are generated by mechanical shearing of DNA, fragments are ligated into plasmids and transformed into bacteria, DNA fragments are sequenced and assembled by computer analysis

what is new generation of DNA sequencing techniques

massively parallel, require no cloning or gels, goal to sequence a genome for under $1000

next generation sequencing applications

targeted sequencing, metagenomics 16S and ITS, genotype by sequencing, RNA sequencing

most important step in next generation sequencing

library prep

what is targeted sequencing

isolation and amplification of a specific region for sequencing, subset of genes of regions of the genome is amplified for sequencing

what is amplicon sequencing

sequencing of specific targets at one time

what does amplicon sequencing allow for

multiple targets at time (multiplex)