Search results
Results From The WOW.Com Content Network
The template DNA must be eliminated by enzymatic digestion with a restriction enzyme such as DpnI, which is specific for methylated DNA. All DNA produced from most Escherichia coli strains would be methylated; the template plasmid that is biosynthesized in E. coli will, therefore, be digested, while the mutated plasmid, which is generated in ...
The number of copies of the gene is then amplified using polymerase chain reaction (PCR). Simultaneously, the same restriction enzymes are used to digest (cut) the destination. The idea behind using the same restriction enzymes is to create complementary sticky ends , which will facilitate ligation later on.
Amplification can be directed across the altered restriction site, and the products digested with the restriction enzyme. This method has been called Cleaved Amplified Polymorphic Sequence (CAPS). Alternatively, the amplified segment can be analyzed by allele-specific oligonucleotide (ASO) probes, a process that can often be done by a simple ...
Restriction digest is most commonly used as part of the process of the molecular cloning of DNA fragment into a vector (such as a cloning vector or an expression vector).The vector typically contains a multiple cloning site where many restriction site may be found, and a foreign piece of DNA may be inserted into the vector by first cutting the restriction sites in the vector as well the DNA ...
Amplified fragment length polymorphism (AFLP-PCR or AFLP) is a PCR-based tool used in genetics research, DNA fingerprinting, and in the practice of genetic engineering. Developed in the early 1990s by Pieter Vos, [ 1 ] AFLP uses restriction enzymes to digest genomic DNA , followed by ligation of adaptors to the sticky ends of the restriction ...
The first few steps of COBRA, and the molecular changes caused by each step to methylated and unmethylated CpG sites. Combined Bisulfite Restriction Analysis (or COBRA) is a molecular biology technique that allows for the sensitive quantification of DNA methylation levels at a specific genomic locus on a DNA sequence in a small sample of genomic DNA. [1]
Uracil N-glycosylase (UNG) is utilized to eliminate carryover polymerase chain reaction (PCR) products in PCR. This method modifies PCR products such that in a new reaction, any residual products from previous PCR amplifications will be digested and prevented from amplifying, but the true DNA templates will be unaffected. [16]
N6-methyladenine (m6A) is the product of the addition of a methyl group (CH 3) at position 6 of the adenine. This modified nucleotide is absent from the vast majority of eukaryotes, with the exception of C. elegans , [ 3 ] but is widespread in bacterial genomes, [ 4 ] as part of the restriction modification or DNA repair systems.