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Chemical methods for decoding cytosine modifications in DNA.

Booth MJ, Raiber EA, Balasubramanian S - Chem. Rev. (2014)

View Article: PubMed Central - PubMed

Affiliation: †Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW United Kingdom.

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and therefore does not introduce a strand cleavage... This results in hydroysis with loss of ammonia... The resulting uracil bisulfite adduct also decreases at pH values below 5.0 due to the protonation of the most effective catalytic species, sulphite... with 5mC than with C, due to inhibition of the adduct formation from the electronic effect of the methyl group... buffers”; however, there has been no definitive examination of their effectiveness... resolution map of 5mC, 5hmC and 5fCs... 5hmC and, through oxidiation of 5mC with the TET enzymes, have been might be feasible to use such nanopores to discriminate 5mC and 5hmC By chemically altering the primary alcohol of 5hmC it is It is now possible to decode 5mC, 5hmC,

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Detection of 5mC and5hmC with restriction endonucleases. 5mC and5hmC can be distinguished at an HpaII/MspI restriction digestion site by comparing digestion reactions. HpaII only digests DNA that does not contain 5mC or 5hmC. MspI digests DNA that contains 5mC and 5hmC. Glucosylationof 5hmC (5gC) inhibits digestion with MspI. 5hmCcan be distinguished by comparing glucosylation/MspI digestion and MspI only digestion. 5mC can bedistinguished by comparing glucosylatin/MspI digestionand HpaII digestion.
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fig6: Detection of 5mC and5hmC with restriction endonucleases. 5mC and5hmC can be distinguished at an HpaII/MspI restriction digestion site by comparing digestion reactions. HpaII only digests DNA that does not contain 5mC or 5hmC. MspI digests DNA that contains 5mC and 5hmC. Glucosylationof 5hmC (5gC) inhibits digestion with MspI. 5hmCcan be distinguished by comparing glucosylation/MspI digestion and MspI only digestion. 5mC can bedistinguished by comparing glucosylatin/MspI digestionand HpaII digestion.

Mentions: The two restriction enzymes regularlyused to detect 5mC are HpaII and MspI, which both cut at the sameDNA sequence; CCGG. This sequence is ideal as it contains a CpG dinucleotide,which is where the majority of 5mC resides in mammals.27HpaII is methylation-sensitiveand will only cut a CCGG sequence that does not contain 5mC, whereas MspI is methylation-insensitive and will cut a CCGG sequencewith or without 5mC (Figure 6).


Chemical methods for decoding cytosine modifications in DNA.

Booth MJ, Raiber EA, Balasubramanian S - Chem. Rev. (2014)

Detection of 5mC and5hmC with restriction endonucleases. 5mC and5hmC can be distinguished at an HpaII/MspI restriction digestion site by comparing digestion reactions. HpaII only digests DNA that does not contain 5mC or 5hmC. MspI digests DNA that contains 5mC and 5hmC. Glucosylationof 5hmC (5gC) inhibits digestion with MspI. 5hmCcan be distinguished by comparing glucosylation/MspI digestion and MspI only digestion. 5mC can bedistinguished by comparing glucosylatin/MspI digestionand HpaII digestion.
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fig6: Detection of 5mC and5hmC with restriction endonucleases. 5mC and5hmC can be distinguished at an HpaII/MspI restriction digestion site by comparing digestion reactions. HpaII only digests DNA that does not contain 5mC or 5hmC. MspI digests DNA that contains 5mC and 5hmC. Glucosylationof 5hmC (5gC) inhibits digestion with MspI. 5hmCcan be distinguished by comparing glucosylation/MspI digestion and MspI only digestion. 5mC can bedistinguished by comparing glucosylatin/MspI digestionand HpaII digestion.
Mentions: The two restriction enzymes regularlyused to detect 5mC are HpaII and MspI, which both cut at the sameDNA sequence; CCGG. This sequence is ideal as it contains a CpG dinucleotide,which is where the majority of 5mC resides in mammals.27HpaII is methylation-sensitiveand will only cut a CCGG sequence that does not contain 5mC, whereas MspI is methylation-insensitive and will cut a CCGG sequencewith or without 5mC (Figure 6).

View Article: PubMed Central - PubMed

Affiliation: †Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW United Kingdom.

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

and therefore does not introduce a strand cleavage... This results in hydroysis with loss of ammonia... The resulting uracil bisulfite adduct also decreases at pH values below 5.0 due to the protonation of the most effective catalytic species, sulphite... with 5mC than with C, due to inhibition of the adduct formation from the electronic effect of the methyl group... buffers”; however, there has been no definitive examination of their effectiveness... resolution map of 5mC, 5hmC and 5fCs... 5hmC and, through oxidiation of 5mC with the TET enzymes, have been might be feasible to use such nanopores to discriminate 5mC and 5hmC By chemically altering the primary alcohol of 5hmC it is It is now possible to decode 5mC, 5hmC,

Show MeSH