Characterization of virus-like particles associated with the human faecal and caecal microbiota.

Characterization of virus-like particles associated with the human faecal and caecal microbiota.

Research in microbiology (2014-12-03)

Lesley Hoyles, Anne L McCartney, Horst Neve, Glenn R Gibson, Jeremy D Sanderson, Knut J Heller, Douwe van Sinderen

ABSTRACT

This work represents an investigation into the presence, abundance and diversity of virus-like particles (VLPs) associated with human faecal and caecal samples. Various methodologies for the recovery of VLPs from faeces were tested and optimized, including successful down-stream processing of such samples for the purpose of an in-depth electron microscopic analysis, pulsed-field gel electrophoresis and efficient DNA recovery. The applicability of the developed VLP characterization method beyond the use of faecal samples was then verified using samples obtained from human caecal fluid.

MATERIALS

Product Number

Brand

Product Description

426148

Sigma-Aldrich

Boric acid-¹¹B, ≥99 atom % ¹¹B

185574

Sigma-Aldrich

1,1,2,2-Tetrabromoethane, 98%

86760

Sigma-Aldrich

1,1,2,2-Tetrabromoethane, purum p.a., for separation of mineral compounds, ≥98.0% (GC)

46067

Sigma-Aldrich

Ethidium bromide solution, suitable for fluorescence, ~1% in H₂O

E1385

Sigma-Aldrich

Ethidium bromide solution, BioReagent, Molecular Biology, 500 μg/mL in H₂O

E1510

Sigma-Aldrich

Ethidium bromide solution, BioReagent, Molecular Biology, 10 mg/mL in H₂O

32900

Supelco

Benalaxyl-M, PESTANAL^®, analytical standard

E8008

Sigma-Aldrich

Ethylenediaminetetraacetic acid solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture

B7660

Sigma-Aldrich

Boric acid, BioXtra, ≥99.5%

B6768

Sigma-Aldrich

Boric acid, BioReagent, Molecular Biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%

B2645

Sigma-Aldrich

Boric acid, tablet, 1 g boric acid per tablet

B7901

Sigma-Aldrich

Boric acid, suitable for electrophoresis, ≥99.5%

E6758

Sigma-Aldrich

Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture

Sigma-Aldrich

Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder

EDS

Sigma-Aldrich

Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)

E7637

Sigma-Aldrich

Ethidium bromide, BioReagent, Molecular Biology, powder

03609

Sigma-Aldrich

Ethylenediaminetetraacetic acid, BioUltra, ≥99.0% (KT)

03620

Sigma-Aldrich

Ethylenediaminetetraacetic acid, ≥98.0% (KT)

03690

Sigma-Aldrich

Ethylenediaminetetraacetic acid disodium salt solution, BioUltra, pH 8.0, ~0.5 M in H₂O

15663

Sigma-Aldrich

Boric acid, BioUltra, Molecular Biology, ≥99.5% (T)

339067

Sigma-Aldrich

Boric acid, 99.97% trace metals basis

431788

Sigma-Aldrich

Ethylenediaminetetraacetic acid, 99.995% trace metals basis

202878

Sigma-Aldrich

Boric acid, 99.999% trace metals basis

E9884

Sigma-Aldrich

Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder

V900106

Sigma-Aldrich

Ethylenediaminetetraacetic acid, Vetec^™, reagent grade, 98%

B0252

Sigma-Aldrich

Boric acid, ReagentPlus^®, ≥99.5%

B0394

Sigma-Aldrich

Boric acid, ACS reagent, ≥99.5%

11607

Sigma-Aldrich

Boric acid, puriss., meets analytical specification of Ph. Eur., BP, NF, 99.5-100.5%, powder

31146

Sigma-Aldrich

Boric acid, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., buffer substance, ≥99.8%

E8751

Sigma-Aldrich

Ethidium bromide, ~95% (HPLC)