Whole Insect DNA Extraction at Intertek

Introduction

This document provides general instructions for sampling whole beetles or other whole insects in 96-format plates for DNA extraction at Intertek after DNA extraction optimization has been completed.

DNA Extraction Optimization - Intertek requests 2-4 plates of dummy (i.e., non-precious and expendable tissue) collected and prepared in a feasible workflow to establish a preferred method to prepare samples for DNA extraction going forward. Things to take into consideration are tissue type, tissue amount, and preservation method.

If tissue types or different sample types other than what was recommended following the DNA extraction optimization procedure with Intertek are desired, discuss the matter in advance with your BI coordinator. Additional instructions may apply.

Communicating with Intertek

Required plasticware

How to prepare and collect samples

1. Design the genotyping experiment and determine the number of plates needed (number of samples/94), striving for complete plates in sets of 4.

  • Best pricing is achieved at 16 plates per order.

  • Consider adding more samples to fill out any partial plates. A full plate is 94 samples.

2. Send an email to bi-genotyping@cornell.edu with your plate count request and a brief summary of your experiment.

3. Create your sample list ahead of time.

  • Before samples are placed in tubes, annotate the sample list with sex or any other information that must be collected

  • A partially-filled Intertek order form is available below.

4. Pre-label all your empty sample plates with human-readable text. Include barcodes if possible.

  • If you use a fixed-well format sample plate, decide on your plate layout before you start sampling.

  • Always leave wells G12 and H12 empty in all plates. They are used by DArT for lab controls during sequencing.

  • Documentation is essential to link the sample ID to the experimental ID.

5. Prepare the sampling tools, sample list, and labeled plates. (Be sure your other logistics are in order before you start sampling.)

6. Keep the samples and plates cool on wet ice while sampling.

7. Select the insects (developmental stage, sex, population, location) to be sampled .

8. Collect samples of the appropriate amount and age, based on your DNA extraction optimization test.

9. Preserve the samples.

10. Seal the plates with Intertek-approved caps or mats.

11. Request BI’s approval to ship plates (if BI is paying for genotyping) by sending an email to bi-genotyping@cornell.edu with the following information:

  • Your completed order form

  • Any information about sample ploidy or wild origin

  • A photograph of your plate labels

Caution. Failure to receive prior approval by BI will cause processing delays, increase cost to you, and may flag your order for destruction.

12. BI will apply the appropriate quote number and PO number. BI will double check that all required details are provided and approve the plates for shipping.

13. Print the completed Intertek order form that you receive back from BI.

14. Insert into an outer package a) the printed order form and b) the sealed plates.

  • The outer package can be a cardboard box.

  • An inner package, such as a styrofoam, box can be used to prevent plate breakage during shipment, but is not required.

  • Do not send plates on dry or wet ice. It is not necessary for fully dried leaf tissues and is more likely to moisten samples rendering them unusable.

15. Ship the entire package to the Intertek lab via courier according to the instructions below “How to send samples”.

16. Email the completed Order Form to Intertek and include the courier and tracking information.

17. Alert Intertek that an order is incoming by emailing a draft order form to Agritech Sweden (agritech.sweden@intertek.com) with bi-genotyping@cornell.edu in the CC field.

How to send samples

1. Intertek recommends the courier DHL for the fastest logistics. However, other couriers can be used.

2. Fill out the Commercial Invoice for your chosen international shipping company.

3. Create a Pro-forma document on your institution’s letterhead with the following statement: “For analytical purposes only, not for breeding, will be destroyed.”

4. Attach the Commercial Invoice, the Pro-forma document, and the shipping manifest to the outside of the parcel.

Caution

  • You MUST write the correct Commodity Code on the Commercial Invoice.

  • You MUST include the statement “For analytical purposes only, not for breeding, will be destroyed” on the Pro-forma document.

  • You MUST attach all paperwork on the outside of the parcel.

5. Send the parcel by courier to the following address:

6. You will receive an order number and confirmation by email when samples have arrived at the destination.

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Breeding Insight is funded by USDA at Cornell University. For more information, please visit http://www.breedinginsight.org.