Manual of Standard Operating Procedures

Purpose

The purpose of this protocol is to facilitate a streamlined process for biospecimen collection, storage, shipping, and processing. By following these protocols, biospecimens will be collected and handled uniformly, ensuring that data acquired from prospective uses are as comparable as possible, regardless of the collection site.

Scope

This manual is broadly broken into four main sections: Collection Kit Overview, Biospecimen Collection, Shipping Instructions, and Quality Control. Biospecimen collection kits are assembled in the INCLUDE DS-Biorepository central lab, located at the University of Colorado Anschutz Medical Campus, and shipped to various collection sites. Using these kits and following the protocols outlined in the Biospecimen Collection section, the collection sites will at a minimum, collect and store tongue swabs, cells for downstream DNA isolation, cells stabilized for RNA isolation, and plasma. In addition to the minimum, these protocols will facilitate the collection of cryopreserved buffy coat, red blood cells, and whole blood fixed for immunophenotyping using Cytometry by time of flight (CyTOF®). Samples will be batch-shipped once a predetermined threshold of 4 participants (U.S. Sites) or 10 participants (international sites) have been collected, or 3 months have passed since the last shipment.

Required Materials

Below is a list of the required materials for biospecimen collection, storage, and return shipment to the central biorepository. Once received by the central lab, these samples will be stored for prospective research use. Many of these materials are provided by the DS-Biorepository central lab as part of the pre-assembled kits. Materials provided in these kits, as well as materials and equipment required but not supplied, are detailed below and in their respective portions of the Biospecimen Collection section.

Material	Source	Vendor	Catalog #
1000 ul Barrier Tips, Sterile	To be provided by collection site
10X Phosphate Buffered Saline (PBS)	Supplement Kit (for Child/Adult Kits)	Rockland	MB-008
200 ul Barrier Tips, Sterile	To be provided by collection site
20 ul Barrier Tips, Sterile	To be provided by collection site
BD PhosflowTM Lyse/Fix Buffer 5x	Supplement Kit (for Child/Adult Kits)	BD Bioscience	558049
Biospecimen Bag, Small	Child, Adult, Tasso Kits	Fisher Scientific	12-888-306
Biospecimen Bag, Medium	Child, Adult, Saliva	Fisher Scientific	23-700-205
Conical tubes, 15 mL	To be provided by collection site
Conical tubes, 50 mL	To be provided by collection site
Cryogenic Vials, 1.2 mL	Child, Adult Kits	Corning	430487
Dimethyl Sulfoxide (DMSO), Sterile Filtered	Supplement Kit (for Child/Adult Kits)	Sigma-Aldrich	D2438-50ML
Dry Ice	To be provided by collection site
Dry Ice Label	Child, Adult Kits	Uline	S-2844
Fetal Bovine Serum-Heat inactivated (FBS)	Supplement Kit (for Child/Adult Kits)
Gauze	Tasso Kits	Fisher Scientific	22-362178
Gloves	To be provided by collection site
Insulated Shipping Box	To be provided by collection site
K2 EDTA Vacutainer, 10 mL	Adult Kits	BD Bioscience	366643
K2 EDTA Vacutainer, 4 mL	Child Kits	BD Bioscience	367861
Liquid Medical Waste Fluid Solidifier	Child, Adult Kits	Ecolab	LTSP3000
Matrix 0.5 mL Screw Top Tubes	Child, Adult, Tasso Kits	Thermo Scientific	3745-BR
Matrix Tube Caps, Colorless	Child, Adult, Tasso Kits	Thermo Scientific	4470
Matrix Tube Caps, Red	Child, Adult Kits	Thermo Scientific	4470RED
Maxpar® Cell Staining Buffer	Supplement Kit (for Child/Adult Kits)	Standard BioTools	201068
Microcentrifuge Tubes, 1.5 mL	To be provided by collection site
Microcentrifuge tubes, 2.0. mL	Tasso, Child, Adult Kits	Thermo Scientific	3463
OMNIgene Tongue Swab	Tasso, Child, Adult Kits	DNA Genotek	OMR-120
ORAgeneTM Saliva Kit	Saliva Kits	DNA Genotek	OGR-600
PAXgene® Blood DNA Vacutainer, 2.5 mL	Child Kits	BD Bioscience	761165
PAXgene® Blood DNA Vacutainer, 8.5 mL	Adult Kits	BD Bioscience	761115
PAXgene® Blood RNA Vacutainer, 2.5 mL	Child, Adult Kits	BD Bioscience	762165
Red Blood Cell Lysis Buffer	Supplement Kit (for Child/Adult Kits)	Thermo Scientific	A1049201
Rigid cardboard box (return of samples)	To be provided by collection site
RNAlaterTM Stabilization Solution	Tasso Kit	Invitrogen	AM7020
Saliva Collection Sponge	Tasso, Child, Adult Kits	DNA Genotek	OG-AC1
Serological Pipets, 10 mL	To be provided by collection site
Styrofoam box (return of samples)	To be provided by collection site
Tasso+® EDTA Kit, RUO	Tasso Kits	Tasso	K1025-01-EDT-G01-RUO
Therapak™ Aqui-Pak™ Absorbent Sleeves	Child, Adult, Tasso, Saliva Kits	Fisher Scientific	22-130-043
UltraPure Distilled Water	Supplement Kit (for Child/Adult Kits)	Invitrogen	10977-015
UN3373 Label	All Kits	Uline	S-12521
Ziploc Bag, 12″ x 15″	All Kits	Fiser Scientific	NC9870745