Sampling Process at Therion

Overview of sample processing:

Samples are received in the lab daily, Monday through Saturday. Packages are sorted by type (blood tubes, blood cards and brushes), opened and the packaging discarded. (We do not routinely gather information from the packaging, so please do not put information there. Put it on the sample submission form.) The information from the sample submission form is entered into the database.

Once 44 samples are received a plate is assembled. It can take from 3 days to 1 week for enough samples to arrive to complete a plate. DNA is extracted from the samples. There are three separate processes used, one for each sample type, which can take 1-3 days to complete. Once all the DNA samples are complete and amplified the plate is loaded onto the genotyping machine, which runs overnight. The following day, the scientist reviews all data from the machine and submits the results back to the office.

A report is generated showing all parentage information for each sample. This report is reviewed by me. Every sample is checked by hand to ensure the results are correct and make sense. Parentage is either confirmed or exclusions are noted. Samples that did not produce results from enough markers are sent back to the lab for a re-run. [Exceptions to this include: brush samples with fewer than 7 markers, blood cards that were received damp, blood cards that have been re-run twice but do not have enough markers and blood tubes that have been re-run three times and do not have sufficient markers. In these cases resubmission of the sample is requested.] Any corrections on the report are entered into the database and DNA profile certificates are prepared. The profiles and the reported data are submitted electronically to the ASCA Office.

Cause of delays:

The major cause for delay in reporting results is re-runs. Members can help avoid this problem by carefully following the sample submission instructions.

1. Many blood cards are received damp, which means too much blood was applied and/or they were not allowed to completely air dry prior to folding shut and mailing. These samples often become contaminated with mold, which compromises the results.

2. Brush samples often do not contain enough cellular material. Saliva is not sufficient for our process. The brush must be applied with enough pressure to the inside of the cheek to scrape cells from the mouth. It is also important not to get food or hair onto the brush.

3. Blood samples are the most robust sample we receive and we can always go back to it and re-isolate the DNA. It is important the blood collection tube has not expired (or if it is expired it can still be used as long as clear anti-coagulant liquid is readily visible in the tube before the blood is added). After the blood is added, the tube must be gently shaken (slowly rocked back and fourth) so the anticoagulant mixes thoroughly with the blood.

Another cause of delay is incomplete information on the sample submission form. Please fill it out completely and correctly. We have gotten entries that say “you know this information”, parentage information entered in the wrong places, etc. It takes time to track down the correct information and is disruptive to sample flow. It is also better to submit multiple sire information with the sample rather than after the sample has been excluded.

Some samples are also held back if the parents were submitted at the same time and their profile is incomplete. We will then review and report once the parents’ results are done.

Occasionally, a multi-sire parentage cannot be confirmed to a single sire. This usually results when the two potential sires are closely related (father and son, brothers, etc.). We can run an extended marker set to try and differentiate the sires. This means all samples (including dam) must be run with the additional markers. Additional fees are charged for this extended testing so owner permission must be obtained first. This testing is also done separately from the regular sample processing. All of these things cause delays for reporting of the associated samples.

Results:

The vast majority of samples proceed through the lab smoothly. Most parentages are verified. There are about one or two sire exclusions a month and very rarely a dam is excluded. Most of these are eventually resolved with more information, by testing another sire or resubmitting samples that may have been labeled incorrectly.

Contacting the lab:

It is best not to contact the lab directly, and we would greatly appreciate it you would call or email the ASCA Office first as they have more information than we do. If you must contact the lab please use email (casna@theriondna.com or gergits@theriondna.com) not the telephone. Include sample name, registry number, owner’s name and parentage information if applicable. We will investigate your question and get back to you as soon as we can.