We use the GenDx NGS HLA typing protocols. The sample requirements for HLA typing projects analyzing the core 11 loci are listed below. Samples that do not meet these requirements will not be rejected. The results from our internal testing with samples that did not meet these requirements produced incomplete or poor data, so these requirements are strictly enforced.
For projects targeting single loci or targeted loci groups, we can accept smaller volumes of gDNA as long as the concentration still meets the requirements. Please contact us if you have any questions about minimum sample volumes or DNA concentration.
|Sample type||Min. volume||Container||Notes|
|Buccal swab||N/A||1 buccal swab in sealed tube||None|
|500uL||1mL - 6mL tubes||Blood collection tubes should use EDTA or ACD as an anticoagulant. Vacutainer K2 EDTA blood collection tubes usually have lavender or pink caps. Collection tubes should NOT contain heparin. We do not recommend using Zymo DNA Shield tubes.|
|Cell pellet||≤ 3mL||1mL - 2mL tubes||5M cells, * See important note below
Suspend in PBS (Phosphate Buffered Saline)
|DNA||≥ 60uL||1mL - 2mL tubes||Concentration ≥ 30ng/uL, Mass >=60ng.
** See important note below
|Nasal swab||N/A||1 nasal swab in sealed tube||None|
|PBMC||200uL||1mL - 2mL tubes||Cell concentration = 5M cells/mL|
|Saliva||1mL||1mL - 2mL tubes||None|
|Tissue||1g||1mL - 2mL tubes||None|
* Cell Pellets: The DNA extraction kit we use for cell pellets requires the sample to contain at least 5M cells in order to yield the minimum required DNA concentration for library prep. If the sample does not contain 5M cells, The Sequencing Center cannot guarantee the extraction will meet the minimum concentration requirement. You may send multiple vials in order to meet the 5M cells requirement.
** DNA: It is essential that the DNA concentration be at least 30ng/uL. HLA results will often be incomplete if the minimum concentration is not met. The Sequencing Center uses and recommends using fluorometric methods for DNA quantitation (i.e. Qubit). If you must use a spectrophotometer for quantitation, please provide samples that exceed the minimum concentration since spectrophotometry often registers higher DNA concentrations than fluorometry.