1) Compare throughputs of various sequencing technologies and discuss the impact
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Question
1) Compare throughputs of various sequencing technologies and discuss the impact of sequencing using a case study taking an example of human or plant disease
2) Design a workflow that integrate various omics such as genomics, transciptomic and metabolomics to understand biology of cancer
3) Briefly comment on application of epigentic workflows to understand unknown causative factors for stress in plants or animals
..ooo C Spire 8:03 AM 1 Compare throughputs of various sequencing technologies and discuss the impact of sequencing 2. Design a workflow that integrate various omics such as genomics,transcrigtomic and 3. Briefly comment on application of epigenetic workfigws to understand unknowm causative using a case study taking a ex of-men or ase factors for stress in plants or animalsExplanation / Answer
Chain termination : Sanger method.
The enzymatic chain termination method developed by Frederick Sanger based on the DNA polymerase dependent synthesis of complementary DNA strand in presence of dNTPs and ddNTPs. The DNA synthesis reaction is randomly terminated whenever a ddNTP is added to the growing oligonucleotide chain, resulting in truncated products of varying length with an appropriate ddNTP at their 3' terminua. The product are separated by size using agarose gel electrophoresis and the terminal ddNTPs are used to reveal the DNA sequence of template strand.
Chemical degradation method : Maxam- Gilbert method
The chemical degradation method introduced by Allan Madam and Walter Gilbert 1977. In this method, the sequence of D's DNA molecule is determined by treatment with chemicals that cut the molecule at specific nucleotide positions.
Pyroseqencing method :
It was developed by Mostafa Ronaghi and Pal Buren 1996 , the addition of dNTPs to the end of growing strand is detectable because it is accompanied by the release of a flash of light.
Next generation sequencing technologies
Illumina sequencing based on principal of sequencing by synthesis
In torrent sequencing based on incorporation of appropriate dNTP and if correct dNTP added PPi and H+ is realeased which changes the pH and detector detects it.
Human genome project which was done by Sanger sequencing helped to identify locus of various gene responsible for genetic diseases.E.g. by using ESTs to determine the gene expressing them and to determine if they have any nucleotide variation or SNPs when compared to normal individual.
Also human genome sequencing made it easy to reaveal mutations and metabolic diseases
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