![]() ![]() ![]() An alternative approach is the use of R loops recognizing proteins as intermediates. To state that the DNA region analyzed forms R loops the mutation rate should increase, should be specific to the non-template ssDNA strand and should be reverted by RNaseH action. Both, the bisulfate anion and AID deaminate cytidines present in ssDNA giving rise to uracil and/or subsequent mutation. ![]() R loops can be inferred in vivo by determining the mutation profile of the DNA produced by sodium bisulfate treatment or by the action of activation-induced cytidine deaminase (AID). In all the cases, the signal observed should disappear after RNaseH treatment. Other direct methods would be DNA:RNA immunoprecipitation (DRIP) or immunofluorescent in yeast chromosomal spreads or human cell with anti-DNA:RNA hybrid antibodies. One direct method is based on the isolation of all nucleic acids from the cell followed by treatment with RNase A and DNase I that leaves only DNA-RNA hybrids intact. R loops can be detected directly by physical and molecular methods or indirectly by genetic approaches. Rondón, in Reference Module in Life Sciences, 2017 Detection of R Loops In Vivo ![]()
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