Targeted Genotyping Specific Requirements

GDNA QUANTITY

As TG products follow fully automated assay processing we require larger gDNA quantity to account for dead volume required for liquid handling robots in addition to volume used for gDNA QC. For samples with gDNA quantity range of 50 to 100ng/µl – 25µl is optimal. Samples with gDNA quantity lower than 50ng/µl and especially lower than 20ng/µl require minimum of 35µl.

We can process the samples with lower volumes (never lower than 15µl) however the assay accuracy will be lower and variability higher.

The sample volume should not exceed 50µl.

Plastic-ware

Due to liquid handling robots requirements gDNA is required to be shipped in two types of fully skirted, V-shape bottom 96 well PCR plates:

  • 1st Choice – Twin-Tec from Eppendorf (Product code: 0030128648) secured with Strips of 8 clear FLAT caps from Sarstedt (Product code 65.1998.400.1)
  • 2nd choice – FrameStar from 4titude (Product code: 4ti-0960/C) secured with Strips of 8 clear FLAT caps from Sarstedt (Product code 65.1998.400.1

FrameStar 4titude plates are the second choice as they require additional care while closing the plate with strip caps. The cap lip can be very easily damaged if strip cap is not perfectly aligned with the plate and pressed down. Damaged caps will leak during transport resulting in loss of samples and potential cross-contamination.

If you are having problems with sourcing the required plastic-ware we can arrange the shipment of it for an additional fee.

Alternatively, we can transfer the gDNA from the received plates to the required plastic-ware before processing the assay; however it will attract additional charge and may result in loss of accuracy if the volume shipped was lower than recommended.

1 Sarstedt discontinued strip caps 65.989.002 (made in Germany) and 65.1998.002 (made in the USA). These are replaced by 65.1998.400 (made in Great Britain or Hungary) these can be used for the strip chains and the Eppendorf twin Tec 96 well PCR plates.

Number of Samples Per Ordered Service

TG services are processed in the format of 384 plate, thus we require the services to be ordered in multiples of four 94 plates (wells G12 and H12 must be empty for the controls).

We can process partial 384 plates, however the cost per sample will increase: the less samples the higher cost.

Positive Control (PC) Requirements

We require minimum 500µl of ~25ng/µl of good quality (HMW present) PC gDNA shipped in EB buffer (or equivalent) in molecular grade tube. If there are no residual nucleases or gDNA shearing reagents in PC prep it can be shipped at room temperature. To check for presence of residual nucleases or gDNA shearing reagents please perform “mock digest” as described here: https://www.diversityarrays.com/faq/

PC can be shipped as precipitated and dried pellet. If selecting this shipment method please include the information of gDNA quantity.

Alternatively, we can process the extraction in our facility. If this is the preferred option please ship few grams of tissue following our recommendations for tissue shipments: https://www.diversityarrays.com/orderinstructions/ with the exception of molecular grade plastic ware which need to be suited to the tissue size.

Please include the printed copy of Positive Control Specification document (download below) and all other required documentation depending what (tissue/gDNA) and from where (domestic/international) is shipped.

Please contact us (link to contact) in case you have any doubts or additional questions.

Page last updated: May 13, 2021, 1:03 pm