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Field Collection and Sampling

Field Collection and Sampling. Starting your collection. What to sample: Tissue What to consider: Age, Season What methodology to used for your biological questions What is the natural h istory of your organism eg . Life history etc.

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Field Collection and Sampling

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  1. Field Collection and Sampling courtesy of Carol Ritland

  2. Starting your collection • What to sample: Tissue • What to consider: Age, Season • What methodology to used for your biological questions • What is the natural history of your organism eg. Life history etc. http://images.google.ca/imgres?imgurl=http://www.christiananswers.net/kids/animals6.jpg&imgrefurl=http://www.christiananswers.net/kids/find2.html&h=375&w=500&sz=70&hl=en&start=5&usg=__d6sSz-0SC0vP_NuogTY5sRsvsxY=&tbnid=a9-2ftNtkcqeBM:&tbnh=98&tbnw=130&prev=/images%3Fq%3Danimals%26gbv%3D2%26hl%3Den%26client%3Dfirefox-a%26rls%3Dorg.mozilla:en-US:official%26sa%3DG

  3. Starting your collection….cont’d • Check for permission and necessary permits for sampling • Check for location of field sites • Check on logistics of field collection http://www.monkeygrove.com/scribbles/2002/PaperWork.gif

  4. Conifer abundance: affect on soil richness Photos = L. Ritland courtesy of Carol Ritland

  5. Starfish abundance: affect on barnacles Photos = L. Ritland

  6. What should you collect? • What could you collect? • Where would you collect? • When would you collect? • 5) How would you collect? • How much would you collect? • 7) Why did you collect them?

  7. Tissue (Always go fresh if possible): • Plant: Seed, Leaves, Flower, Pollen, Bark, Xylem, Roots • Animal: Reproductive tissue, muscle, skin, hair, scat, blood, ear/toe/tail clip, fin, tooth, sloughed skin, saliva • Fungal: Hyphae, spores, fruiting bodies • Bacterial: single isolate culture • Destructive vs Non Destructive methods

  8. Factors to consider when collecting: • Age: • Plant = actively growing material such as apical points, seedlings • Animal = actively dividing tissue (buccal and blood cells) • Fungus = young fruiting bodies (pure culture) • Bacteria = liquid culture • Season: • For conifer = use early spring bud burst especially for DNA markers

  9. How to sample: • Sampling schemes: • Linear • Quadratic squares • Distance between samples • For animals: migration, reproductive strategies, life cycle • For plants: clonality, roots, reproductive strategies, life cycle • Consult a statistician? (Gene expression studies) courtesy of Carol Ritland

  10. Sampling cont’d • Tools for sampling: • Ideally flash freezing samples with liquid nitrogen and transportation under ultra low temperature • Clean and if possible sterilize collection tools between samples • Pack samples with foil or proper containers eg. cryovials • Label all samples with non water based ink and protect with clear tape/paper and pencil courtesy of Carol Ritland

  11. Liquid nitrogen vapour tanks courtesy of Carol Ritland

  12. Ultralow temp. Liq N2 tanks • Pending on size • Ultralow temp can last from 10 days to 3 weeks • Can be used on aircrafts courtesy of Carol Ritland

  13. Use sterile technique when sampling • Keep meticulous records • Identify any problems in the field for given sample • Do not depend on your memory alone courtesy of Carol Ritland

  14. Long Term Storage: • Ideally all tissues should be kept at ultra low temperature (minus 70 to 80°C) • Certain tissues (blood, animal tissues cut into small pieces <1mm2 ) could be stored in 100% ethanol and saturated EDTA • Plants (small amounts) could be desiccated with lots of silica beads with lots of changes of beads courtesy of Carol Ritland

  15. No No..s • Items to avoid: • Unnecessary chemicals eg. Formalin • historical samples that has been treated with fixatives • degraded samples • freezing and thawing of tissue • freezer burn (improper storage conditions) • improper inventory of samples courtesy of Carol Ritland

  16. More to ponder….. • Items to consider: • For certain molecular marker (eg. AFLP, microarray) use the same tissue type (developmental differences could cause error when genotyping) • If possible collect all samples within a season over a same span of time (eg. For microarray analysis) • Collect more samples than required for pilot study and lost of samples courtesy of Carol Ritland

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