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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]
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 ...
Nucleic acids are often denatured by including urea in the buffer, while proteins are denatured using sodium dodecyl sulfate, usually as part of the SDS-PAGE process. For full denaturation of proteins, it is also necessary to reduce the covalent disulfide bonds that stabilize their tertiary and quaternary structure, a
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]
DNA polymerase epsilon proves to be best suited for nucleotide excision repair. DNA polymerase epsilon is independent of both PCNA and RFC, and produces mostly ligated DNA products. It is also found that under one condition where DNA polymerase epsilon require PCNA and RFC: nucleotide excision repair in the presence of single strand binding ...
Before annealing can occur, one of the strands may need to be phosphorylated by an enzyme such as kinase to allow proper hydrogen bonding to occur. The term annealing is often used to describe the binding of a DNA probe , or the binding of a primer to a DNA strand during a polymerase chain reaction .
Three more DNA polymerases have been found in E. coli, including DNA polymerase III (discovered in the 1970s) and DNA polymerases IV and V (discovered in 1999). [9] From 1983 on, DNA polymerases have been used in the polymerase chain reaction (PCR), and from 1988 thermostable DNA polymerases were used instead, as they do not need to be added in ...
DNA polymerase I also has 3' to 5' and 5' to 3' exonuclease activity, which is used in editing and proofreading DNA for errors. The 3' to 5' can only remove one mononucleotide at a time, and the 5' to 3' activity can remove mononucleotides or up to 10 nucleotides at a time.