Using PCR, you want to amplify an approximately 1 kbexon of the human autosomal gene encoding theenzyme phenylalanine hydroxylase from the genomicDNA of a patient suffering from the autosomal recessive condition phenylketonuria (PKU).a. Why might you wish to perform this PCR amplification in the first place, given that the sequence ofthe human genome has already been determined?b. Calculate the number of template molecules thatare present if you set up a PCR reaction using1 nanogram (1 × 10−9 grams) of chromosomalDNA from blood cells as the template. Assume thateach haploid genome contains only a single genefor phenylalanine hydroxylase and that the molecular weight of a base pair is 660 grams per mole. Thehaploid human genome contains 3 × 109base pairs.c. Calculate the number of PCR product moleculesyou will obtain if you perform 25 PCR cycles andthe yield from each cycle is exactly twice that ofthe previous cycle. What would be the mass ofthese PCR products taken together?
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.
Using PCR, you want to amplify an approximately 1 kb
exon of the human autosomal gene encoding the
enzyme phenylalanine hydroxylase from the genomic
DNA of a patient suffering from the autosomal recessive condition phenylketonuria (PKU).
a. Why might you wish to perform this PCR amplification in the first place, given that the sequence of
the human genome has already been determined?
b. Calculate the number of template molecules that
are present if you set up a PCR reaction using
1 nanogram (1 × 10−9 grams) of chromosomal
DNA from blood cells as the template. Assume that
each haploid genome contains only a single gene
for phenylalanine hydroxylase and that the molecular weight of a base pair is 660 grams per mole. The
haploid human genome contains 3 × 109
base pairs.
c. Calculate the number of PCR product molecules
you will obtain if you perform 25 PCR cycles and
the yield from each cycle is exactly twice that of
the previous cycle. What would be the mass of
these PCR products taken together?
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