New method boosts the study of regulation of gene activity BCMHouston nature
Sequences of all oligonucleotides used in this study are available at Additional file 6: Table S5. 6mA modified and unmodified control oligos were synthesized at GeneLink, Inc. All other oligos, including 5mC modified oligo , were synthesized at Sigma. 4mC modified oligo was generated by treating dsDNA oligo with BamHI methyltransferase according to the manufacturer’s instructions .
Microbial community standard DNA was purchased from Zymo Research . One microgram of genomic DNA was first fragmentized to 100–300 bp using Covaris S220 Focused-ultrasonicator. Fragmented gDNA was ligated to TruSeq adaptor using NEBNext Ultra™ II DNA Library Prep Kit for Illumina . Unmodified control DNA was made by amplifying adaptor-ligated DNA using Q5 DNA polymerase . Both native and amplified DNA was first annealed to excessive protective adaptor sequences in 5 mM adaptor oligos and 0.
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