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Solution hybrid selection capture for the recovery of functional full-length eukaryotic cDNAs from complex environmental samples.

Bragalini C, Ribière C, Parisot N, Vallon L, Prudent E, Peyretaillade E, Girlanda M, Peyret P, Marmeisse R, Luis P - DNA Res. (2014)

Bottom Line: After two successive rounds of capture, >90% of the resulting cDNAs were GH11 sequences, of which 70% (38 among 53 sequenced genes) were full length.Sequencing of polymerase chain reaction-amplified GH11 gene fragments from the captured sequences highlighted hundreds of phylogenetically diverse sequences that were not yet described, in public databases.This protocol offers the possibility of performing exhaustive exploration of eukaryotic gene families within microbial communities thriving in any type of environment.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences and Systems Biology, University of Turin, viale Mattioli 25, Turin 10125, Italy Ecologie Microbienne, UMR CNRS 5557, USC INRA 1364, Université de Lyon, Université Lyon 1, Villeurbanne 69622, France.

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Electrophoretic separation of cDNAs obtained following two consecutive solution hybridization selection. Captured cDNAs from the four soil samples PUE, BRH, BRE and BEW were run on an Agilent DNA 12000 microfluidic chip. Each band could encompass one or several unique but abundant GH11 cDNAs.
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DSU030F1: Electrophoretic separation of cDNAs obtained following two consecutive solution hybridization selection. Captured cDNAs from the four soil samples PUE, BRH, BRE and BEW were run on an Agilent DNA 12000 microfluidic chip. Each band could encompass one or several unique but abundant GH11 cDNAs.

Mentions: SHS was performed on cDNAs synthesized from polyadenylated mRNAs extracted from four different forest soils. Electrophoregrams of all cDNAs recovered after two successive rounds of capture were characterized by a background smear of which emerged discrete bands ranging in size from 300 to 1,500 bp (Fig. 1).Figure 1.


Solution hybrid selection capture for the recovery of functional full-length eukaryotic cDNAs from complex environmental samples.

Bragalini C, Ribière C, Parisot N, Vallon L, Prudent E, Peyretaillade E, Girlanda M, Peyret P, Marmeisse R, Luis P - DNA Res. (2014)

Electrophoretic separation of cDNAs obtained following two consecutive solution hybridization selection. Captured cDNAs from the four soil samples PUE, BRH, BRE and BEW were run on an Agilent DNA 12000 microfluidic chip. Each band could encompass one or several unique but abundant GH11 cDNAs.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263301&req=5

DSU030F1: Electrophoretic separation of cDNAs obtained following two consecutive solution hybridization selection. Captured cDNAs from the four soil samples PUE, BRH, BRE and BEW were run on an Agilent DNA 12000 microfluidic chip. Each band could encompass one or several unique but abundant GH11 cDNAs.
Mentions: SHS was performed on cDNAs synthesized from polyadenylated mRNAs extracted from four different forest soils. Electrophoregrams of all cDNAs recovered after two successive rounds of capture were characterized by a background smear of which emerged discrete bands ranging in size from 300 to 1,500 bp (Fig. 1).Figure 1.

Bottom Line: After two successive rounds of capture, >90% of the resulting cDNAs were GH11 sequences, of which 70% (38 among 53 sequenced genes) were full length.Sequencing of polymerase chain reaction-amplified GH11 gene fragments from the captured sequences highlighted hundreds of phylogenetically diverse sequences that were not yet described, in public databases.This protocol offers the possibility of performing exhaustive exploration of eukaryotic gene families within microbial communities thriving in any type of environment.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences and Systems Biology, University of Turin, viale Mattioli 25, Turin 10125, Italy Ecologie Microbienne, UMR CNRS 5557, USC INRA 1364, Université de Lyon, Université Lyon 1, Villeurbanne 69622, France.

Show MeSH
Related in: MedlinePlus