The short DNA shown below is to be sequenced. Using your knowledge of how the Sanger method works, in the gel diagram, draw in the bands that will appear when DNA polymerase is added to the reaction along with the four different nucleotide mixtures indicated. Note that some of these mixtures are not what would normally be used in a sequencing reaction. Dideoxwnucleotides (ddNTPs) are added in relatively small amounts. The asterisk represents a radioactive label. *5' --- 3'-OH 3' --- ACGACGCAGGACATTAGAC-5' Nucleotide mixtures: A. DATP, dTTP, ACTP, ¿GTP, ddTIR (given) B. DATR, dTTP, ¿CTP, AGTP, ddATP C. dTTP, dGTP, dCTP. ddCTP, ddATR D. DATP, ¿CTP, dTTP, ddGTP А В С D |LL ||
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.
This scenario based on DNA sequencing. DNA sequencing is the process of determining the order of nucleotides in DNA. DNA sequencing in various type such as
- Maxam and Gilbert method.
- Chain termination method. or Sanger sequencing,
- semiautomated method.
- automated method.
- Pyrosequencing.
- The whole-genome shotgun sequencing method.
- Clone by the clone sequencing method.
- Next-generation sequencing method. etc.
Sanger sequencing is a method for determining the nucleotide sequence of DNA, that involves electrophoresis and is based on the random incorporation of chain-terminating dideoxynucleotides.
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