
A DNA sequencing reaction was performed with fragment 5−XXXGCGATCGYYYY−3′ as the template, dideoxy GTP, all the four dNTPs, and the required primers and enzymes. XXXX and YYYY in the given DNA fragment represent primer binding sites. The set of fragments obtained during the reaction will be (the primers are not shown in the amplified fragments).
(a) 5'-CGATCGC-3' only
(b) 5-CG-3, 5-CGCTAG-3', 5'-CGCTAGC-3'
(c) 5'-CG-3, 5'-CGATCG-3, 5'-CGATCGC-3'
(d) 5-G-3, 5'-GCG-3, 5'-GCGATCG-3'
Answer
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Hint: It's the method of determining the sequence of nucleotides (As, Ts, Cs, and Gs) during a piece of DNA. Sequencing a whole genome (all of an organism’s DNA) remains a whole task. Here the breaking of DNA occurs resulting in numerous pieces that are sequenced and then gathered into a group called "consensus."
Complete answer
A DNA sequencing response was performed with the part 5−XXXGCGATCGYYYY−3′ on the grounds that the template, dideoxy GTP, all the four dNTPs, and consequently the necessary primers and enzymes. XXXX and YYYY inside the given DNA part speak to primer binding sites. The arrangement of parts acquired during the response will be CGATCGC since, in such a case that the primer is connected to the XXXX restricting site, its polymerization starts and its complementary fragment is CGATCGC.
Additional information
In Sanger sequencing, the DNA sample to be sequenced is combined during a tube with primer, DNA polymerase, and DNA nucleotides (dATP, dTTP, dGTP, and dCTP). The four dye-labeled, chain-terminating dideoxynucleotides are added also, but in much smaller amounts than the standard nucleotides.
The mixture is first heated to denature the template DNA (separate the strands), then cooled in order that the primer can bind to the single-stranded template. When the primer has bound, the temperature is raised once more, permitting DNA polymerase to synthesize new DNA going from the primer. DNA polymerase will continue adding nucleotides to the chain until it happens to feature a dideoxynucleotide rather than a traditional one. At that time, no further nucleotides are often added, therefore the strand will end with the dideoxynucleotide.
So the correct answer is ‘5'-CGATCGC-3' only’.
Note: Sanger sequencing gives high-quality sequences for relatively long stretches of DNA (up to about 900 base pairs). It typically won’t sequence individual pieces of DNA, like bacterial plasmids or DNA copied in PCR.
However, Sanger sequencing is dear and inefficient for larger-scale projects, like the sequencing of a whole genome or metagenome.
Complete answer
A DNA sequencing response was performed with the part 5−XXXGCGATCGYYYY−3′ on the grounds that the template, dideoxy GTP, all the four dNTPs, and consequently the necessary primers and enzymes. XXXX and YYYY inside the given DNA part speak to primer binding sites. The arrangement of parts acquired during the response will be CGATCGC since, in such a case that the primer is connected to the XXXX restricting site, its polymerization starts and its complementary fragment is CGATCGC.
Additional information
In Sanger sequencing, the DNA sample to be sequenced is combined during a tube with primer, DNA polymerase, and DNA nucleotides (dATP, dTTP, dGTP, and dCTP). The four dye-labeled, chain-terminating dideoxynucleotides are added also, but in much smaller amounts than the standard nucleotides.
The mixture is first heated to denature the template DNA (separate the strands), then cooled in order that the primer can bind to the single-stranded template. When the primer has bound, the temperature is raised once more, permitting DNA polymerase to synthesize new DNA going from the primer. DNA polymerase will continue adding nucleotides to the chain until it happens to feature a dideoxynucleotide rather than a traditional one. At that time, no further nucleotides are often added, therefore the strand will end with the dideoxynucleotide.
So the correct answer is ‘5'-CGATCGC-3' only’.
Note: Sanger sequencing gives high-quality sequences for relatively long stretches of DNA (up to about 900 base pairs). It typically won’t sequence individual pieces of DNA, like bacterial plasmids or DNA copied in PCR.
However, Sanger sequencing is dear and inefficient for larger-scale projects, like the sequencing of a whole genome or metagenome.
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