PMID- 28986786
OWN - NLM
STAT- MEDLINE
DCOM- 20180611
LR  - 20180820
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1654
DP  - 2017
TI  - DNA Methylation Profiling Using Long-Read Single Molecule Real-Time Bisulfite 
      Sequencing (SMRT-BS).
PG  - 125-134
LID - 10.1007/978-1-4939-7231-9_8 [doi]
AB  - For the past two decades, bisulfite sequencing has been a widely used method for 
      quantitative CpG methylation detection of genomic DNA. Coupled with PCR amplicon 
      cloning, bisulfite Sanger sequencing allows for allele-specific CpG methylation 
      assessment; however, its time-consuming protocol and inability to multiplex has 
      recently been overcome by next-generation bisulfite sequencing techniques. 
      Although high-throughput sequencing platforms have enabled greater accuracy in 
      CpG methylation quantitation as a result of increased bisulfite sequencing depth, 
      most common sequencing platforms generate reads that are similar in length to the 
      typical bisulfite PCR size range (~300-500 bp). Using the Pacific Biosciences 
      (PacBio) sequencing platform, we developed single molecule real-time bisulfite 
      sequencing (SMRT-BS), which is an accurate targeted CpG methylation analysis 
      method capable of a high degree of multiplexing and long read lengths. SMRT-BS is 
      reproducible and was found to be concordant with other lower throughput 
      quantitative CpG methylation methods. Moreover, the ability to sequence up to 
      ~1.5-2.0 kb amplicons, when coupled with an optimized bisulfite-conversion 
      protocol, allows for more thorough assessment of CpG islands and increases the 
      capacity for studying the relationship between single nucleotide variants and 
      allele-specific CpG methylation.
FAU - Yang, Yao
AU  - Yang Y
AD  - Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount 
      Sinai, One Gustave L. Levy Place, Box 1498, New York, NY, 10029, USA. 
      yao.yang@mssm.edu.
FAU - Scott, Stuart A
AU  - Scott SA
AD  - Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount 
      Sinai, One Gustave L. Levy Place, Box 1497, New York, NY, 10029, USA. 
      stuart.scott@mssm.edu.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Sulfites)
RN  - OJ9787WBLU (hydrogen sulfite)
SB  - IM
MH  - CpG Islands/genetics
MH  - DNA Methylation/*genetics
MH  - High-Throughput Nucleotide Sequencing/methods
MH  - Humans
MH  - Nanotechnology/methods
MH  - Polymerase Chain Reaction
MH  - Sequence Analysis, DNA/methods
MH  - Sulfites/*chemistry
OTO - NOTNLM
OT  - Bisulfite sequencing
OT  - CpG islands
OT  - DNA methylation
OT  - Long-read sequencing
OT  - Multiplex DNA methylation analysis
OT  - PacBio sequencing
OT  - SMRT sequencing
EDAT- 2017/10/08 06:00
MHDA- 2018/06/12 06:00
CRDT- 2017/10/08 06:00
PHST- 2017/10/08 06:00 [entrez]
PHST- 2017/10/08 06:00 [pubmed]
PHST- 2018/06/12 06:00 [medline]
AID - 10.1007/978-1-4939-7231-9_8 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2017;1654:125-134. doi: 10.1007/978-1-4939-7231-9_8.