In both methods, the DNA was digested by HpaII (a methylation-sensitive enzyme) and MspI (a methylation-insensitive isoschizomer
enzyme) and analyzed by real-time PCR amplification.
To interpret the H3 motif, primers CV140F and CV120R were designed to introduce a FokI site (an isoschizomer
of BstF5I) in the amplified product by substituting the restriction recognition sequence GGATG for T (nucleotide -147) and G (nucleotide -110), respectively.
16) that incorporated a hot-start PCR and isoschizomer
of the restriction enzyme for more efficient digestion.
The amplified samples were checked for purity and quantity on a standard agarose gel (10 g/L) and digested with Cfol, an isoschizomer
tuoliensis was digested with isoschizomers
Msp I or Hpa II (mixture of EcoR I), the ability to digest the sequence CpCpGpG as influenced by their methylation state.
Thereafter, the genomic DNA was digested with the isoschizomers
HpaII and MspI.
Among the topics are detecting changes in global genome methylation using the cytosine-extension assay, isoschizomers
and amplified fragment length polymorphism for detecting specific cytosine methylation changes, northern blotting techniques for small RNAs, cloning new small RNA sequences, cDNA libraries for virus-induced gene silencing, and detecting and quantifying DNA strand breaks using the random oligonucleotide primed synthesis (ROPS) assay.