You have been given the DNA sequence for a particular fragment of DNA. You then isolated the mRNA made from that DNA and amplified it by PCR. You then determined the sequence of the cDNA obtained from different cells. You notice a difference. In the sequence obtained from DNA sequencing you see that a codon is 5' CAG however on the cDNA sequence it is TAG. These results are confirmed by repeated DNA sequence analysis using DNA and cDNA from different cell cultures (same species and tissue samples). What can explain this? a.The DNA must have been mutated in all the cells that were used to isolate mRNA since the cDNA should always match the genomic sequence. b.Any cDNA made through RT-PCR will have T's substitued for genomic C's that are methyulated. c.The mRNA must have been deaminated at the cytosine. d.The cDNA generated most likely had a technical mistake caused by poor fidelity of the Taq enzyme.
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.
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You have been given the DNA sequence for a particular fragment of DNA. You then isolated the mRNA made from that DNA and amplified it by PCR. You then determined the sequence of the cDNA obtained from different cells. You notice a difference. In the sequence obtained from DNA sequencing you see that a codon is 5' CAG however on the cDNA sequence it is TAG. These results are confirmed by repeated DNA sequence analysis using DNA and cDNA from different cell cultures (same species and tissue samples). What can explain this?
a.The DNA must have been mutated in all the cells that were used to isolate mRNA since the cDNA should always match the genomic sequence.
b.Any cDNA made through RT-PCR will have T's substitued for genomic C's that are methyulated.
c.The mRNA must have been deaminated at the cytosine.
d.The cDNA generated most likely had a technical mistake caused by poor fidelity of the Taq enzyme.
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