PMID- 24060113
OWN - NLM
STAT- MEDLINE
DCOM- 20140407
LR  - 20130924
IS  - 1557-7988 (Electronic)
IS  - 0076-6879 (Linking)
VI  - 531
DP  - 2013
TI  - In-solution fluorescence in situ hybridization and fluorescence-activated cell 
      sorting for single cell and population genome recovery.
PG  - 3-19
LID - B978-0-12-407863-5.00001-0 [pii]
LID - 10.1016/B978-0-12-407863-5.00001-0 [doi]
AB  - Over the past decade, technological advances in whole genome amplification, 
      microfluidics, flow sorting, and high-throughput sequencing have led to the 
      development of single-cell genomics. Single-cell genomic approaches are typically 
      applied to anonymous microbial cells with only morphology providing clues to 
      their identity. However, targeted separation of microorganisms based on 
      phylogenetic markers, such as the 16S rRNA gene, is beginning to emerge in the 
      single-cell genomics field. Here, we describe an in-solution fluorescence in situ 
      hybridization (FISH) protocol which can be combined with fluorescence-activated 
      cell sorting (FACS) for separation of single cells or populations of interest 
      from environmental samples. Sequencing of DNA obtained from sorted cells can be 
      used for the recovery of draft quality genomes, and when performed in parallel 
      with deep metagenomics, can be used to validate and further scaffold metagenomic 
      assemblies. We illustrate in this chapter the feasibility of this FISH-FACS 
      approach by describing the targeted recovery of a novel anaerobic methanotrophic 
      archaeon.
CI  - (c) 2013 Elsevier Inc. All rights reserved.
FAU - Haroon, Mohamed F
AU  - Haroon MF
AD  - Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences, 
      The University of Queensland, St. Lucia, Queensland, Australia.
FAU - Skennerton, Connor T
AU  - Skennerton CT
FAU - Steen, Jason A
AU  - Steen JA
FAU - Lachner, Nancy
AU  - Lachner N
FAU - Hugenholtz, Philip
AU  - Hugenholtz P
FAU - Tyson, Gene W
AU  - Tyson GW
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Enzymol
JT  - Methods in enzymology
JID - 0212271
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Archaea/genetics
MH  - *Flow Cytometry
MH  - *Genetics, Population
MH  - Genome
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Metagenomics/*methods
MH  - Microfluidics/methods
MH  - RNA, Ribosomal, 16S
MH  - Sequence Analysis, DNA
MH  - Single-Cell Analysis
OTO - NOTNLM
OT  - Anaerobic methanotrophic archaea
OT  - Anaerobic oxidation of methane
OT  - FISH-FACS
OT  - Fixation-free FISH
OT  - Fluorescence in situ hybridization
OT  - Fluorescence-activated cell sorting
OT  - In-solution FISH
OT  - Metagenomics
OT  - Single-cell genomics
OT  - Single-population genomics
EDAT- 2013/09/26 06:00
MHDA- 2014/04/08 06:00
CRDT- 2013/09/25 06:00
PHST- 2013/09/25 06:00 [entrez]
PHST- 2013/09/26 06:00 [pubmed]
PHST- 2014/04/08 06:00 [medline]
AID - B978-0-12-407863-5.00001-0 [pii]
AID - 10.1016/B978-0-12-407863-5.00001-0 [doi]
PST - ppublish
SO  - Methods Enzymol. 2013;531:3-19. doi: 10.1016/B978-0-12-407863-5.00001-0.