Journal: Canadian Journal of Veterinary Research

Article Title: Development of an immunocapture-polymerase chain reaction assay using IgY to detect Mycobacterium avium subsp. paratuberculosis

doi:

Figure Lengend Snippet: Determination of Beads A volume required for immunocapture of American Type Culture Collection (ATCC) strain 19698 of Mycobacterium avium subsp. paratuberculosis (MAP). For each Beads A volume used, DNA was run undiluted (N) and at dilutions of 1:10 (−1) and 1:100 (−2). The bead volumes were as follows: lanes 1 to 3, 10 μL; lanes 4 to 6, 20 μL; lanes 7 to 9, 30 μL; lanes 10 to 12, 40 μL. Lane −C — negative water control; lane M — 1-kb molecular weight marker; bp — base pairs.

Article Snippet: The concentrations of 3 field and 2 reference strains of MAP [11992, EA4146, 2945, American Type Culture Collection (ATCC) 12258, and ATCC 19698] were adjusted to 2 × 10 7 cells/mL.

Techniques: Control, Molecular Weight, Marker

Journal: Canadian Journal of Veterinary Research

Article Title: Development of an immunocapture-polymerase chain reaction assay using IgY to detect Mycobacterium avium subsp. paratuberculosis

doi:

Figure Lengend Snippet: Polymerase chain reaction (PCR) results with different concentrations of DNA extracted from ATCC 19698 MAP cells immunocaptured by 40 μL of Beads A incubated at room temperature (RT) for different times; DNA was run undiluted (N) and at dilutions of 1:10 (−1) and 1:100 (−2). The incubation times were as follows: lanes 1 to 3, 15 min; lanes 4 to 6, 30 min; lanes 7 to 9, 60 min. Lane +C — positive-control DNA (ATCC 19698); lane +IC — positive internal-control DNA; lane −C — negative water control; lane M — 1-kb molecular weight marker.

Article Snippet: The concentrations of 3 field and 2 reference strains of MAP [11992, EA4146, 2945, American Type Culture Collection (ATCC) 12258, and ATCC 19698] were adjusted to 2 × 10 7 cells/mL.

Techniques: Polymerase Chain Reaction, Incubation, Positive Control, Control, Molecular Weight, Marker

Journal: Canadian Journal of Veterinary Research

Article Title: Development of an immunocapture-polymerase chain reaction assay using IgY to detect Mycobacterium avium subsp. paratuberculosis

doi:

Figure Lengend Snippet: Results of PCR with the use of 40 μL of Beads A to immunocapture MAP ATCC 19698 spiked in bovine feces (2 × 104 cells/g) incubated at RT for 15 min. Lanes 1 and 2 — DNA diluted 1:10 and 1:100; lanes 3 and 4 — negative-control bovine feces with DNA diluted 1:10 and 1:100; lane 5 — positive-control DNA (ATCC 19698); lane 6 — negative water control; lane M — 1-kb molecular weight marker.

Article Snippet: The concentrations of 3 field and 2 reference strains of MAP [11992, EA4146, 2945, American Type Culture Collection (ATCC) 12258, and ATCC 19698] were adjusted to 2 × 10 7 cells/mL.

Techniques: Incubation, Negative Control, Positive Control, Control, Molecular Weight, Marker

Journal: Canadian Journal of Veterinary Research

Article Title: Development of an immunocapture-polymerase chain reaction assay using IgY to detect Mycobacterium avium subsp. paratuberculosis

doi:

Figure Lengend Snippet: Determination of MagaCell-IgY beads volume required for immunocapture of MAP ATCC 19698 spiked in bovine feces (2 × 104 cells/g) and incubated at RT for 15 min. For each MagaCell-IgY beads volume used, DNA was run undiluted (N) and at dilutions of 1:10 (−1) and 1:100 (−2). The bead volumes were as follows: lanes 1 to 3, 5 μL; lanes 4 to 6, 10 μL; lanes 7 to 9, 15 μL; lanes 10 to 12, 20 μL. Lane M — 1-kb molecular weight marker; lane +C — positive-control DNA (ATCC 19698); lane −C — negative water control.

Article Snippet: The concentrations of 3 field and 2 reference strains of MAP [11992, EA4146, 2945, American Type Culture Collection (ATCC) 12258, and ATCC 19698] were adjusted to 2 × 10 7 cells/mL.

Techniques: Incubation, Molecular Weight, Marker, Positive Control, Control

Journal: Canadian Journal of Veterinary Research

Article Title: Development of an immunocapture-polymerase chain reaction assay using IgY to detect Mycobacterium avium subsp. paratuberculosis

doi:

Figure Lengend Snippet: Results of PCR with the use of 10 μL of MagaCell-IgY beads to immunocapture MAP strain ATCC 19698 spiked in bovine feces at concentrations of 2 × 103 cells/g (lanes 1 to 3), 2 × 104 cells/g (lanes 4 to 6), and 2 × 105 cells/g (lanes 7 to 9). End-point titration was performed with extracted DNA run undiluted (N) and at dilutions of 1:10 (−1) and 1:100 (−2). Lane +C — positive-control DNA (ATCC 19698); lane +IC — positive internal-control DNA; lane −C — negative water control; lane M — 1-kb molecular weight marker.

Article Snippet: The concentrations of 3 field and 2 reference strains of MAP [11992, EA4146, 2945, American Type Culture Collection (ATCC) 12258, and ATCC 19698] were adjusted to 2 × 10 7 cells/mL.

Techniques: Titration, Positive Control, Control, Molecular Weight, Marker

Journal: BMC Research Notes

Article Title: Purification of native HBHA from Mycobacterium avium subsp . paratuberculosis

doi: 10.1186/1756-0500-6-55

Figure Lengend Snippet: Sequence alignment of different HBHA proteins. ( A ) Multiple sequence alignment was performed using the Clustal W program with the BLOSUM64 matrice allowing gaps (−). * indicate identical residues. : indicate conserved substitutions. ; indicate semi-conserved substitutions. Mtb, M. tuberculosis ; BCG, M. bovis BCG; Mah , M. avium subsp . hominissuis and Map , M. avium subsp . paratuberculosis ; Msmeg, M smegmatis . ( B ) Distribution of charges within the C-terminal domain of HBHA. The isoelectric point of each protein is indicated in brackets. Color code: Polar charged residues HKR (Pink); Polar residues GNQSTY (Green); Polar negative residues DE (Blue) and other residues (Red).

Article Snippet: Mycobacterium avium subsp. paratuberculosis (Map) strain K-10 (ATCC BAA-968) was grown at 37°C in Sauton medium supplemented with 2 mg L -1 of mycobactin J (IDVET, Montpellier, France).

Techniques: Sequencing

Journal: Genes & Development

Article Title: Probing the initiation and effector phases of the somatic piRNA pathway in Drosophila

doi: 10.1101/gad.1968110

Figure Lengend Snippet: Transposons, piRNAs, and piRNA precursors in zucchini and squash mutant ovaries. (A) Transcripts of gypsy, idefix, and ZAM transposons were detected by qPCR. (B) Individual piRNAs targeting gypsy and idefix were detected by qPCR. (C) Piwi protein levels in mutant and heterozygous ovary extracts were measured by Western blotting. Tubulin serves as loading control. Celera sequencing strain (S-strain) is shown in addition. (D) Three ∼100-nt regions of flamenco that are normally highly processed into piRNAs were detected by qPCR. The positions of these segments are indicated in Figure 4. qPCR data were normalized to internal controls rp49 (A,C) or bantam (B). Fold changes relative to heterozygous siblings are shown on a linear scale. Error bars represent one standard deviation over three technical replicates.

Article Snippet: Celera sequencing strain (S-strain) is shown in addition. ( D ) Three ∼100-nt regions of flamenco that are normally highly processed into piRNAs were detected by qPCR.

Techniques: Mutagenesis, Western Blot, Sequencing, Standard Deviation