Skip to content
Login Register

#sequencing

2 public questions tagged with this topic.

PCR

Different strains of virus can be identified by

Different viral strains show minimal morphological difference yet distinct genomic sequences. Polymerase chain reaction amplifies target nucleic acids using strain-specific primers, followed by sequencing or probe hybridization to identify mutations, genotypes, and lineages with high sensitivity. Fluorescence and electron microscopy reveal localization or ultrastructure but cannot resolve nucleotide variations. Clinical symptoms overlap among strains and are influenced by host immunity. Therefore, sequence-based amplification provides definitive, rapid, and sensitive discrimination, widely applied for influenza, SARS-CoV-2, and other viruses even at very low viral titers.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

View Details
Transcript analysis

Which of the following is false regarding RNA-seq?

RNA sequencing involves isolation of total RNA or poly-A selected RNA, fragmentation, conversion to complementary DNA by reverse transcription, adapter ligation, and high-throughput sequencing. It provides unbiased transcriptome coverage, enabling quantification of expression, discovery of novel transcripts, detection of alternative splicing isoforms, fusion genes, and allelic expression. Ribosomal RNA is typically depleted to avoid dominating reads, not exclusively sequenced. A library exclusively containing ribosomal RNA would defeat the purpose of profiling messenger RNAs and regulatory non-coding RNAs, hence the statement limiting RNA-seq to ribosomal RNA is fundamentally incorrect.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

View Details