Methods and Techniques in Molecular Biology

Running head:TECHNOLOGY OF MOLECULAR BIOLOGY
TECHNOLOGY OF MOLECULAR BIOLOGY
Name of the student
Name of the university
Author note

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TECHNOLOGY OF MOLECULAR BIOLOGY
Part I
Answer 1.
A and B are identified as a third-generation sequencer, or long-read sequencer and C
is identified as short read sequencer. Long read sequencing is more advantageous than short-
range sequencing. Pacific Biosciences’ (PacBio) single-molecule real-time (SMRT)
sequencing and Oxford Nanopore Technologies’ (ONT) nanopore sequencing are two
important technologies that dominate long-read sequencing. These technologies provide
reads more than 10kb. The data of third-generation sequencing highly differs from second-
generation sequencing. Illumina’s Novaseq, HiSeq and MiSeq instrument produce short-read
sequences that produce up to 600 bases. Short read sequencing is precise, worthy and assisted
by a variety of tools that helps in analysis.
SMRT sequencers detect fluorescence related events that tie in with an inclusion of one
particular nucleotide with the help of polymerase which is fastened to the base of tiny well
whereas nanopore sequencers estimate the ionic current fluctuations when a single-stranded
nucleic acid move through biological nanopores (Nakano et al. 2017). The full-length
transcripts are produced by Iso-Seq method using SMRT sequencing. The Nanopore
sequencing from 500 bp to 2.3 Mb along with 10-30 kb genomic libraries provides the
longest read length (Byrne et al. 2017). In SMRT sequencing, the read length is restricted by
the longevity of the polymerase. The library inserts size range from 250bp to 50kbp. Whole
Genome Sequencing, Epigenetic, targeted sequencing, RNA sequencing and Complex
Population can be done with SMRT Sequencing with high accuracy. On the other hand, in
short-read sequencing natural nucleic acid polymer span eight orders of magnitude in length
and sequence them into small amplified fragments which complicate the task of rebuilding
and evaluating the original molecules whereas the long-read sequencing improves mapping

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TECHNOLOGY OF MOLECULAR BIOLOGY
certainty, detection of structural variants, de novo assembly and transcript isoform
identification (Shi et al. 2017).
The schematic diagram is identified as whole-genome sequencing. The whole-genome
sequencing is considered as the analysis of the entire genomic sequence of a cell at a time,
thus provides a comprehensive characterization of the genome. This process of whole-
genome sequencing will help in detecting single nucleotide variants, large structural variants,
and change in copy number, insertions and deletions (Belkadi et al. 2015). This information
will help to obtain inherited disorders, tracking inherited disorders. The purpose of
conducting a whole genome sequence is it provides a high-resolution and screened base-by-
base. It has the potential to capture both large and small variants and detects potential
causative variants for further study of gene expression and regulation mechanisms.
Additionally, it provides a large volume of data in a short period. The large whole-genome
sequencing provides information for population genetics and is highly used to research any
disease. The small whole-genome sequencing is useful to do molecular epidemiology studies,
environmental metagenomics and infectious disease surveillance. The scaffolds gives data on
the long-range of genomic structure instead of indicating the actual DNA sequence that is
present within the gaps in middle of the contigs. The scaffolding process provides
information on the relative position of genomic segments along the chromosome. The
histogram from Illumina will provide the information on the separations in middle of natively
generated paired-end reads (Gökkaya 2020), the length of reads is generated by PacBio and
Oxford Nanopore technology and the optical maps provide information on the lengths of the
fragments which is mapped by BioNano nanocoding technology (Mikheyev and Tin 2014).
From the alignment of the contigs, the order and orientation are inferred. The long reads line
up to the terminal side of the contig implies their adjacency, and the optical maps infer the