Question

A) Outline the steps and features of Sanger DNA sequencing and compare with Next Generation Sequencing (NGS) methods

B) Why inbreeding is of concern for endangered species and strategies to mitigate potential problems?

Answer 1

Answer

Sanger sequencing is chain termination based method, in which labelled dideoxynuleotides are added and template is allowed for polymerisation. The addition of dideoxynucleotide will terminate the amplification as it lacks 3’-OH group; a pre-requisite for polymerisaton. This product is visualised on agarose gel. Whereas Nextgeneration sequencing is also sanger based technology.

Comparison of sanger sequencing with Next Generation Sequencing (NGS) methods

• NGS is very quick and efficient
• Read length is high
• Read clearity is high

Inbreading is mazor concern for endangered species, as it will give chance to express recessive traits which will reduce their fittest for survival.