In a clinical context, a scientist is working with a viral DNA which is about 24000bps long. There are two known variants of the virus that share almost the same DNA but for a final fragment; with reference to Figure Q2b, the regions A and B are conserved in both variants, while the region C differs and is either 320bps (variant 1) or 380bps (variant 2). The scientist wants to set up a procedure to identify the variant they are dealing with. Viral dsDNA (i) (ii) (iii) Stable region (A) Variable region (C) Figure Q2b Known sequence (B) 5-GACCTCAATGTCCAGCGGTACGCTCATAAA-3' 3'-CTGGAGTTACAGGTCGCCATGCGAGTATTT-5' The scientists want to design a primer to amplify the variable region and to do so, they sequence a small fragment (sequence B) dthe conserved region close to the variable region C. Why is the scientist targeting a region outside of the fragment of interest? The sequence of the fragment B is reported in Figure Q2b. Suggest a primer that can efficiently target this region and duplicate the variable fragment using PCR. Comment on the minimum length your primer should have to be reasonably specific. The scientists were not able identify a suitable stable region to design a reverse primer against and they decided to perform PCR with just one forward primer (using an excess of it), expecting a slower (linear) amplification instead of the normal exponential growth. Explain why this would occur
Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.
![In a clinical context, a scientist is working with a viral DNA which is about 24000bps
long. There are two known variants of the virus that share almost the same DNA but
for a final fragment; with reference to Figure Q2b, the regions A and B are conserved
in both variants, while the region C differs and is either 320bps (variant 1) or 380bps
(variant 2). The scientist wants to set up a procedure to identify the variant they are
dealing with.
Viral dsDNA
(i)
(ii)
(iii)
Stable region (A)
Variable region (C)
Figure Q2b
Known sequence (B)
5-GACCTCAATGTCCAGCGGTACGCTCATAAA-3'
3'-CTGGAGTTACAGGTCGCCATGCGAGTATTT-5'
The scientists want to design a primer to amplify the variable region and to do
so, they sequence a small fragment (sequence B) the conserved region close
to the variable region C. Why is the scientist targeting a region outside of the
fragment of interest?
[3]
The sequence of the fragment B is reported in Figure Q2b. Suggest a primer
that can efficiently target this region and duplicate the variable fragment using
PCR. Comment on the minimum length your primer should have to be
reasonably specific.
The scientists were not able to identify a suitable stable region to design a
reverse primer against and they decided to perform PCR with just one forward
primer (using an excess of it), expecting a slower (linear) amplification instead
of the normal exponential growth. Explain why this would occur](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Fcc012420-4d67-48a3-8e85-bac83b5d2ff3%2F881f2728-bad4-446d-aab1-5b4db68f7668%2Fwu98z0a_processed.jpeg&w=3840&q=75)
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