Cryptosporidium Genotyping for Source

Tracking

Improving our ability to manage the safety of our water supplies.

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Source Tracking Made Easier!

An all in one kit containing everything you need to genotype your Cryptosporidium samples!

Kit contains

1. Slide extraction reagents

2. Simple safe DNA isolation and Purification reagents

3. Primary, Secondary, and Platinum PCR SuperMix.

4. Restriction Enzymes (SspI, VspI, DdeI, plus buffers)

5. C. parvum PCR-RFLP Positive Control Template.

6. Simple to use standardised Electrophoresis reagents with 2% and 4% buffer free E-gel casettes.

7. I-Gel PowerBase (Part of kits 740-05 and 740-06).

8. Dynabeads anti-Cryptosporidium kit (Part of kit 740-05).

9. Spot on Slides (Part of kit 740-05).

10.L10 tubes (Part of kit 740-05).

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BASIC PROTOCOL

50 L of water was passed through a Filta-max™ filter. Processed parasite enumeration using Method 1623 Immunomagnetic separation (IMS) carried out using

Dynabeads-GC Combo Kit

Oocysts enumerated using immunofluorescent antibodies (IFA),

PCR-RFLP analysis. Xiao et al. DNA extracted Ruecker et al All positive reactions digested with Ssp I, Vsp I and Dde I Fractionated by 2% agarose gel electrophoresis and visualized

PCR products excised from the electrophoresis gel, Purified and sequenced (if necessary)

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FIG. 2. Molecular forensic profiling of Cryptosporidium parasites by repetitive nested PCR-RFLP analysis using microscope slides containing three water samples processed by USEPA method 1623.

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FIG. 2. Molecular forensic profiling of Cryptosporidium parasites by repetitive nested PCR-RFLP analysis using microscope slides containing three water samples processed by USEPA method 1623.

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Simple DNA Extraction protocol, no solvents or alcohols

All reagents included

Pre-poured Agarose gel cassettes and standardised buffer-less electrophoresis running system

Call off charts for fragment sizes/patterns