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DNA polymerase, the main enzyme to catalyze the polymerization of free deoxyribonucleotides into a newly forming DNA strand, plays a significant role in the occurrence of this mutation. When DNA polymerase encounters a direct repeat, it can undergo a replication slippage. [4] Strand slippage may also occur during the DNA synthesis step of DNA ...
Excessive magnesium concentrations also stabilize double stranded DNA and prevent complete denaturation of the DNA during PCR reducing the product yield. [ 6 ] [ 7 ] Inadequate thawing of MgCl 2 may result in the formation of concentration gradients within the magnesium chloride solution supplied with the DNA polymerase and also contributes to ...
The process of DNA denaturation can be used to analyze some aspects of DNA. Because cytosine / guanine base-pairing is generally stronger than adenine / thymine base-pairing, the amount of cytosine and guanine in a genome is called its GC-content and can be estimated by measuring the temperature at which the genomic DNA melts. [2]
DNA denaturation occurs when hydrogen bonds between base pairs are disturbed. The non-covalent interactions between antiparallel strands in DNA can be broken in order to "open" the double helix when biologically important mechanisms such as DNA replication, transcription, DNA repair or protein binding are set to occur. [ 19 ]
The polymerase chain reaction is the most widely used method for in vitro DNA amplification for purposes of molecular biology and biomedical research. [1] This process involves the separation of the double-stranded DNA in high heat into single strands (the denaturation step, typically achieved at 95–97 °C), annealing of the primers to the single stranded DNA (the annealing step) and copying ...
In the less extensive technique of equilibrium unfolding, the fractions of folded and unfolded molecules (denoted as and , respectively) are measured as the solution conditions are gradually changed from those favoring the native state to those favoring the unfolded state, e.g., by adding a denaturant such as guanidinium hydrochloride or urea.
The dye nucleotide to be used will likely occur by treatment with a phosphorylation enzyme and biotinylation and reaction of the biotinylated substance with the dye. It is possible that immediate reaction with the dye may also occur, but extending the arm is claimed to increase efficiency in the case of using a mutant form of DNA polymerase. [5]
This is because G-C base pairing have 3 hydrogen bonds between them while A-T base pairs have only 2. DNA with mutations from either A or T to either C or G will create a higher melting temperature. The information also gives vital clues to a molecule's mode of interaction with DNA.