IRRI SERVICE LABORATORIES
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4.B Submit samples to outsourced service providers
Send DNA samples
1. Prepare sample quality and quantity
- Place DNA samples in water when preparing.
- Prepare the concentration and volume of samples according to the number of SNPs. You may refer to the table below.
Caution: Do not fill your 96 wells with more than 120 ul, and your 384 wells with more than 40 ul.
2. Plate the samples
- Place the samples in a 96 well, full-skirted PCR plates. You may use either the FrameStar 96 Well Plate by 4titude or twin.tec® microbiology PCR Plate 96 by Eppendorf.
- Leave wells G12 and H12 blank for controls.
- Seal the plates with Peel Seal by 4titude.
Send leaf samples
1. Select quality samples
- Collect leaf samples that are from the same development stage.
- Remove any live insects, soil, and dirt on the leaf before punching.
- During punching, take samples from the distal end of leaf and away from the main nerve or mid-rib.
2. Prepare sample quantity
- Clean and prepare the sampling device. These can be scissors, single-hole punchers, or semi-automated punchers (e.g. AK-EP100).
- For each sample, punch 2 leaf disks with a diameter of 5-6mm.
Caution: Do not over sample! Too much tissue will prohibit thorough drying & grinding. This may negatively affect the quality of the SNP results.
3. Plate the samples
- Spray the top part of a round-bottom, 96 well, 1.2mL AbGene Storage Plate with 70% ethanol.
- Air-dry the plate using a PlantTrak deionizer to prepare the placement of samples.
- Leave some wells blank for controls. Refer to the table below for the blanks associated per service partner.
4. If samples are missing, physically mark the empty well and make a notation on the form/documents.
5. Seal the plates using a sealing mat (e.g. AB0674).
Label the samples
Label the plates with either human readable text or barcodes. Printed labels are preferred, however you may place labels using adhesive tape and permanent markers Ensure that the plate names match the ones indicated in the submission forms. Additionally, they should meet the following criteria below.
- 1-30 characters (Agriplex)
- 1-21 characters (Intertek)
- No spaces, only underscore or dash
- Place labels on the east side of the plate
Treat samples for shipping
To ensure that the quality of samples are maintained, samples should be freeze dried prior to shipping. If freeze drying is not available, you can add desiccants in generous amounts (e.g. 10 silica gels : 1 leaf punch).
- Place the sealed plates in durable, see-through, and sealable plastic containers.
- Double-bag the plates to ensure their protection during shipping.
- Place the bagged samples into a final container. This can be a box or plastic.
- Secure the final container with either tape, rubber bands, or clips.
- For Intertek samples, label the final container with the scientific name of the crop.