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In biochemistry, denaturation is a process in which proteins or nucleic acids lose folded structure present in their native state due to various factors, including application of some external stress or compound, such as a strong acid or base, a concentrated inorganic salt, an organic solvent (e.g., alcohol or chloroform), agitation and radiation, or heat. [3]
The most famous example is the hyperchromicity of DNA that occurs when the DNA duplex is denatured. [1] The UV absorption is increased when the two single DNA strands are being separated, either by heat or by addition of denaturant or by increasing the pH level. The opposite, a decrease of absorbance is called hypochromicity.
Common lysis buffers contain sodium hydroxide (NaOH) and sodium dodecyl sulfate (SDS). Cell lysis is best done at a pH range of 11.5–12.5. Cell lysis is best done at a pH range of 11.5–12.5. Although simple, it is a slow process, taking anywhere from 6 to 12 hours.
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 ]
Sodium hydroxide, also known as lye and caustic soda, [1] [2] is an inorganic compound with the formula NaOH.It is a white solid ionic compound consisting of sodium cations Na + and hydroxide anions OH −.
Under neutral conditions, hydrogen bonds reform between complementary base pairs, joining single strands to double-stranded DNA. Because the plasmid was so tightly coiled and small before the alkaline conditions were established, it can easily reanneal. The chromosomal DNA, however, because of its lengthy strands, does not anneal. [1]
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 ...
DNA damage inhibits M-CDKs which are a key component of progression into mitosis. In all eukaryotic cells, ATR and ATM are protein kinases that detect DNA damage. They bind to DNA damaged sites and activate Chk1, Chk2, and, in animal cells, p53. Together, these proteins make up the DNA damage response system.