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Next Generation Sequencing

Submitted by rmmcdonald on Mon, 09/30/2019 - 21:06

Through out the past few decade various ways to sequence the genome had been invented. Originally the genome could only be sequenced in small portions and required mostly human oversight/work to set up the sequencing. This method of sequencing was time consuming and expensive. Today, next generation sequencing has revolutionized the way genomes are sequenced. Mass Parallel squencing is acomplished by sonicating a genome and ligating short, universal primers into the cut DNA strands. That is then poured over a "Chip" where individual fragments of DNA bind to unique positions by the primer that was just ligated on. Then synthesis begins on all the segments. A fluorescent ddNTP base pairs and is captured by a special camera. Next the ddNTP is altered so that the 3' end has an OH group where a new ddNTP molecule can bind. This process is repeated over and over again until all the strands are sequenced. Next, a program will run to match all the overlapping segments into one strand. It is convention to have atleast 30 overlapping sequences in one position to maintain accuracy of the sequencing. 30 overlapping segments would be known as having a depth of 30.