DNA SEQUENCING

Frederick Sanger

discovered DNA Sequencing in 1977

1980

the year Fredrick Anger received his Nobel Prize for developing methods for DNA Sequencing

DNA fragmentation - Library preparation - Amplification - Sequencing - Bioinformatics and Data analysis

General steps for Next generation Sequencing

DNA fragmentation

break the targeted DNA into short segments usually 100-300 bp in length (e.g., mechanical methods, enzymatic digestion)

Library preparation

break the targeted adaptors

Amplification

bridge PCR or emulsion PCR

Sequencing

matrix (e.g., Illumina uses flow cells; Ion Torrent uses chips)

Bioinformatics and Data Analysis

process involving base calling, read alignment and variant identification

Whole genome sequencing

• Almost all the nucleotide in the genome including chromosomal DNA and mitochondrial DNA are examined

• Used more often in Research and less common in clinical settings

Exome Sequencing

Entire coding region of an organism is analyzed

RNA Sequencing

Assembly of RNA transcripts including mRNA, rRNA, tRNA, microRNA, and noncoding RNA are studied

Targeted Sequencing

Commonly used for cancer patients, interrogates hundreds of targeted genes

1977

What year that DNA Sequencing was found?