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: At a 1:100 dilution of DNA, the IC was amplified in both cases but the MAP gene was not amplified. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 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).

Techniques: 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: At a 1:100 dilution of DNA, the IC was amplified in both cases but the MAP gene was not amplified. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 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).

Techniques: Polymerase Chain Reaction, Incubation, Positive 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: At a 1:100 dilution of DNA, the IC was amplified in both cases but the MAP gene was not amplified. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 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).

Techniques: Incubation, Negative Control, Positive 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: At a 1:100 dilution of DNA, the IC was amplified in both cases but the MAP gene was not amplified. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 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).

Techniques: Incubation, Molecular Weight, Marker, Positive 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: At a 1:100 dilution of DNA, the IC was amplified in both cases but the MAP gene was not amplified. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 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).

Techniques: Titration, Positive Control, Molecular Weight, Marker