What is ligation based sequencing?

What is ligation based sequencing?

Sequencing by ligation is a DNA sequencing method that uses the enzyme DNA ligase to identify the nucleotide present at a given position in a DNA sequence. Instead, the mismatch sensitivity of a DNA ligase enzyme is used to determine the underlying sequence of the target DNA molecule.

Which sequencing method uses sequencing by ligation method?

Life Technologies instruments such as the ABI SOLiD System™ utilize sequencing by ligation instead of DNA polymerase extension. Sequencing by ligation involves the use of multiple primers offset by one base at the 3′ end of the adapter.

How does SMRT sequencing work?

At the heart of SMRT Sequencing is the SMRT Cell, which contains millions of tiny wells called zero-mode waveguides (ZMWs). Single molecules of DNA are immobilized in these wells, and as the polymerase incorporates each nucleotide, light is emitted, and nucleotide incorporation is measured in real time.

What are examples of next generation sequencing?

NGS can be used to sequence entire genomes or constrained to specific areas of interest, including all 22 000 coding genes (a whole exome) or small numbers of individual genes. Example of next generation sequencing (NGS) raw data-BRAF V600E mutation in melanoma.

What is ABI SOLiD sequencing?

ABI SOLiD: Sequencing by ligation technology is marketed by Applied Biosystems, USA. The name SOLiD stands for Small Oligonucleotide Ligation and Detection System. This technology was developed by George Church in 2005, and was further improved and distributed by Applied Biosystems in 2007 (Voelkerding et al., 2009).

What is sequencing by ligation?

Sequencing by ligation is a DNA sequencing method that harnesses the mismatch sensitivity of DNA ligase to determine the underlying sequence of nucleotides in a given DNA sequence (Ho et al., 2011).

How accurate is SOLiD 3plus DNA ligation?

For example, the base at read position 5 is assayed by primer number 2 in ligation cycle 2 and by primer number 3 in ligation cycle 1. According to ABI, the SOLiD 3plus platform yields 60 gigabases of usable DNA data per run. Due to the two base encoding system, an inherent accuracy check is built into the technology and offers 99.94% accuracy.

How does SOLiD sequencing work?

During sequencing, each base in the template is sequenced twice, and the resulting data are decoded according to this scheme. SOLiD (Sequencing by Oligonucleotide Ligation and Detection) is a next-generation DNA sequencing technology developed by Life Technologies and has been commercially available since 2006.