What is the minimum sample amount for DIA proteomics?

For DIA proteomics, the minimum depends on the tissue type. Normal animal tissues: 5 mg minimum. Fatty tissue/skin: 100 mg minimum. Serum/plasma without enrichment: as little as 1 μl.

How many biological replicates are required?

A minimum of 3 biological replicates is required. For animal studies, 3–6 replicates recommended. For clinical samples, 6–10 replicates recommended.

What if my sample type is not listed?

If your sample type is not listed, please contact MetwareBio. Our scientific team will evaluate your sample and provide specific guidance.

Sample Requirements - MetwareBio

Q: How much plasma or serum do I need for untargeted metabolomics?

For untargeted metabolomics, the recommended amount is 100 μl per sample, with a minimum of 20 μl. For targeted panels (e.g., steroid hormones, oxylipins), you may need 50–100 μl. Always provide extra volume if possible.

Biomedical Sample Requirements for Metabolomics

Biological duplicates: A minimum of 3 replicates is required; 3–6 replicates for animal samples; 6–10 for clinical samples.

Biomedical Sample Classification

Liquid I Common

Plasma, Serum, Hemolymph, Whole Blood, Milk, Egg White

Liquid II

CSF, Interstitial Fluid, Uterine Fluid, Pancreatic Juice, Bile, Pleural Effusion, Follicular Fluid, Fallopian Tube Fluid, Postmortem Fluid, Culture Medium, Fermentation Broth, Tears, Bone Marrow (liquid)

Liquid III

Seminal Plasma, Amniotic Fluid, Prostatic Fluid, Rumen Fluid, Respiratory Condensate, Gastric Lavage, BALF, Urine, Sweat, Saliva, Sputum

Tissue I Common

Small Animal Tissues, Placenta, Blood Clot, Mycelium, Nematode, Zebrafish (whole fish), Bone Marrow (solid), Nail

Tissue II

Large Animal Tissues, Whole Insect Body, Wings, Pupa, Eggs, Large Fungi, Cartilage, Bone (solid)

Tissue III

Zebrafish Organs, Insect Organs, Whole Microinsect Body (e.g., Drosophila)

Solid I

Feces, Intestinal Contents, Lyophilized Fecal Powder

Solid II

Milk Powder, Microbial Fermentation Product (solid), Culture Medium (solid), Lyophilized Tissue/Plant/Egg Powder

Solid III

Honey, Nasal Mucus, Sputum, Fresh Egg Yolk

Solid IV

Sludge, Soil

Cell I Common

Adherent Cells, Animal Cell Lines

Cell II

E. Coli, Yeast Cells

Cell III

Small Amount of Fungal Mycelial Balls, Cyanobacteria, Large Amount of Bacteria, Slime Mold

Organelle I

Lysosomes, Mitochondria, Endoplasmic Reticulum

Organelle II

Exosomes, Extracellular Vesicles

Special Samples

Skin Tape/Patch, Test Strips, Swab, Hair

Sample Amount Requirements

Sample Class	Recommended	Minimum	Untargeted (Plus)	TM Widely Targeted	Quant. Lipidomics
Liquid I	100 μl	20 μl	✓	✓	✓
Liquid II	100 μl	20 μl	✓	✓	✓
Liquid III	500 μl	50 μl	✓	✓	✓
Tissue I	100 mg	20 mg	✓	✓	✓
Tissue II	500 mg	20 mg	✓	✓	✓
Tissue III	20 units	/	✓	✓	✓
Solid I	200 mg	20 mg	✓	✓	✓
Solid II	100 mg	20 mg	✓	✓	✓
Solid III	100 mg	20 mg	✓	✓	✓
Solid IV	600 mg	300 mg	✓	✓	✓
Cell I	1×10⁶	5×10⁵	✓	✓	✓
Cell II	1×10¹⁰	5×10⁸	✓	✓	✓
Cell III	100 mg	/	✓	✓	✓
Organelle I	4×10⁷ cells / 0.2g	1×10⁷ cells / 0.1g	✓	✓	✓
Organelle II	2×10⁹ / 40μg protein	1×10⁹ / 20μg protein	✓	✓	✓
Special I–IV	1–2 pieces	1 piece	✓	✓	✓

Sample Class	Recommended	Minimum	Energy Metabolism	Tryptophan	One-Carbon	Amino Acids	Organic Acids
Liquid I	100 μl	20 μl	✓	✓	✓	✓	✓
Liquid II	100 μl	20 μl	50–100 μl	✓	✓	✓	✓
Liquid III	500 μl	50 μl	✓	✓	✓	✓	✓
Tissue I/II	100–500 mg	20 mg	50–100 mg	✓	✓	✓	✓
Solid I/II	100–200 mg	20 mg	50–200 mg	✓	✓	✓	✓
Solid III	100 mg	20 mg	0.5–2 g	0.5–2 g	0.5–2 g	0.5–2 g	0.5–2 g
Solid IV	600 mg	300 mg	✓	✓	✓	✓	✓
Cell I	1×10⁶	5×10⁵	✓	✓	✓	✓	✓
Cell II	1×10¹⁰	5×10⁸	✓	✓	✓	✓	✓
Organelle I/II	varies	varies	✓	✓	✓	✓	✓

Sample Class	Bile Acids	Neurotransmitters	Steroid Hormones	Oxylipins	SCFAs	FFAs	Sugars
Liquid I	✓	✓	50–100 μl	50–100 μl	50–100 μl	50–100 μl	✓
Liquid II	✓	50–100 μl	50–100 μl	≥100 μl	50–100 μl	50–100 μl	✓
Liquid III	✓	✓	✓	✓	✓	✓	✓
Tissue I/II	✓	50–100 mg	✓	✓	✓	✓	✓
Solid I–III	✓	50–200 mg	0.5–2 g	0.5–2 g	0.5–2 g	0.5–2 g	0.5–2 g
Solid IV	✓	200–600 mg	✓	✓	✓	✓	600 mg
Cell I	✓	✓	✓	✓	1×10⁷–2×10⁷	✓	5×10⁶–1×10⁷
Cell II	✓	✓	✓	✓	✓	✓	✓

Plant Sample Requirements for Metabolomics

Plant duplicates: A minimum of 3 replicates is required.

Plant Sample Classification

Tissue Common

Root, Stem, Leaf, Fruit, Flower, Bud, Node, Callus, Seed

Liquid I

Root Exudate

Liquid II

Wine

Liquid III

Juice, Fermentation Broth

Liquid IV

Oils, Essential Oils, Honey, Nectar, Paste

Liquid V

Herb Extract, Herb Decoction, Plant Tissue Fluid, Phloem Sap, Leaf Intercellular Fluid

Cell

Fungal Mycelial Balls/Mycelium, Unicellular Algae, Bacteria (Pellet), Slime Mold

Organelle

Plant-derived Exosome-like Nanoparticles

Solid I

Lyophilized Plant Powder

Solid II

Sludge, Soil

Plant Sample Amount Requirements

Sample Class	Recommended	Minimum	Widely Targeted / Flavonoid	Quant. Lipidomics	Plant Hormones	Carotenoid	Anthocyanin	Sugars	Org. Acids / AA	Energy	SCFAs	FFAs
Tissue	600 mg	300 mg	✓	✓	100–300 mg	✓	✓	✓	✓	200–300 mg	200–300 mg	200–300 mg
Liquid I	10 ml	3 ml	✓	200–500 μl	2 mL	2 mL	2 mL	2 mL	2 mL	100 μl–2 mL	2 mL
Liquid II	5 ml	50 μl	✓	200–500 μl	2 mL	2 mL	2 mL	2 mL	2 mL	100 μl–2 mL	2 mL
Liquid III	5 ml	50 μl	✓	200–500 μl	50–200 μl	100–500 μl	100–500 μl	100–500 μl	100–200 μl	50–200 μl	50–200 μl
Liquid IV	500 μl	100 μl	✓	10–100 mg	50–200 μl	100–500 μl	100–500 μl	100–500 μl	100–200 μl	50–200 μl	50–200 μl
Liquid V	500 μl	200 μl	✓	✓	50–200 μl	100–500 μl	100–500 μl	100–500 μl	100–200 μl	50–200 μl	50–200 μl
Cell	100 mg	/	✓	20–100 mg	/	500–600 mg	/	✓	✓	✓	✓	✓
Organelle	2×10⁹ / 40μg	1×10⁹ / 20μg	✓	✓	/	/	/	/	/	/	/	/
Solid I	600 mg	200 mg	✓	✓	/	✓	✓	20–100 mg	50–100 mg	50–100 mg	20–100 mg	20–100 mg
Solid II	600 mg	300 mg	✓	/	/	/	/	600 mg	✓	✓	✓	✓

Sample Requirements for Proteomics

Sample Type	DIA / DDA		Label-free Peptidomics		Phosphorylation		Ubi/Ac/Suc		Lactylation		N-Glycosylation
Sample Type	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.
Normal Tissues ▼ Heart, Liver, Spleen, Lung, Kidney, Brain, Intestine, Stomach, Pancreas, Gallbladder, Testis, Ovary, Breast, Prostate, Thyroid, Adrenal Gland, Lymph, Muscle, Blood Vessel	20 mg	5 mg	100 mg	50 mg	30 mg	15 mg	100 mg	50 mg	150 mg	75 mg	25 mg	15 mg
Fatty Tissue, Skin, Synovium	200 mg	100 mg	1 g	500 mg	300 mg	150 mg	/	/	/	/	/	/
Bone, Shell, Eggshell	1 g	500 mg	1 g	500 mg	/	/	/	/	/	/	/	/
Cartilage Tissue	400 mg	150 mg	1 g	500 mg	/	/	/	/	/	/	/	/
Hair	500 mg	200 mg	1 g	500 mg	/	/	/	/	/	/	/	/
Red Bone Marrow	20 mg	5 mg	/	/	30 mg	15 mg	100 mg	50 mg	150 mg	75 mg	30 mg	15 mg
Yellow Bone Marrow	50 mg	20 mg	/	/	50 mg	30 mg	200 mg	100 mg	200 mg	100 mg	50 mg	30 mg
Soft-bodied Insects ▼ Fruit Flies, etc.	30 mg	5 mg	/	/	30 mg	15 mg	100 mg	50 mg	150 mg	75 mg	30 mg	15 mg

Sample Type	DIA / DDA		Label-free		Phosphorylation		Ubi/Ac/Suc		Lactylation		N-Glyco
Sample Type	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.
Young Tissue (Leaves, Petals, Callus)	50 mg	20 mg	200 mg	100 mg	200 mg	100 mg	1 g	500 mg	1 g	500 mg	100 mg	50 mg
Mature Tissue, Stems, Algae, Fungi	500 mg	200 mg	2 g	1.5 g	500 mg	300 mg	2 g	1 g	2 g	1 g	250 mg	150 mg
Bark, Roots, Fruits, Tubers	1 g	500 mg	2 g	1.5 g	3 g	1.5 g	5 g	3 g	5 g	3 g	1.5 g	1 g
Pollen, Anthers	40 mg	15 mg	/	/	/	/	/	/	/	/	/	/
Lyophilized Plant Powder	300 mg	150 mg	/	/	/	/	/	/	/	/	/	/

Sample Type	DIA / DDA		Label-free		Phosphorylation		Ubi/Ac/Suc		Lactylation		N-Glyco
Sample Type	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.	Rec.	Min.
Primary Cells	3×10⁶	1×10⁶	1×10⁷	5×10⁶	2×10⁷	1×10⁷	2×10⁷	2×10⁷	3×10⁷	1.5×10⁷	2×10⁷	1×10⁷
Serum/Plasma (no enrichment)	10 μl	1 μl	200 μl	50 μl	/	/	/	/	/	/	50 μl	10 μl
Urine	5 ml	2 ml	20 ml	10 ml	/	/	/	/	/	/	/	/
Bacteria	100 mg	50 mg	300 mg	150 mg	300 mg	150 mg	500 mg	200 mg	500 mg	200 mg	150 mg	75 mg
FFPE Section	15 slides (5–10 μm)		/	/	/	/	/	/	/	/	/	/
Extracted Protein	100 μg	50 μg	/	/	500 μg	200 μg	5 mg	3 mg	6 mg	4 mg	500 μg	250 μg

Sample Requirements for DIA Proteomics: IP, Pull-down, and SDS-PAGE Derived Samples

Technology	Acceptable Sample Types	Sample Requirements	Our Intake	MS-compatibility Notes	Recommended Controls
IP / Co-IP / Pull-down	Bead–Ab/bait–target complexes (bead pellet)	Submit 20 μL bead pellets. Perform IP/PD from ~1×10⁷ cells. After final wash, rinse beads with PBS, remove excess liquid, ship on dry ice.	On-bead digestion → LC-MS/MS	Indicate the antibody used (host/isotype/clone) or pull-down bait format.	Isotype IgG, beads-only, Input/Flow-through
IP / Co-IP / Pull-down	Eluates (acid/peptide/competitive/denaturing)	Submit ≤50 μL eluate with ~10–20 μg total protein. Use mild MS-compatible elution buffer. Do NOT use Laemmli buffer.	Buffer exchange → digestion → LC-MS/MS	Specify elution buffer composition. If high SDS, we perform acetone precipitation.	Bait-only resin, free-ligand competition
SDS-PAGE	Gel bands (clean excised pieces)	Target band(s) clearly visible. Load ≥10 μg/lane. Provide gel image with MW marker. Ship with icepacks.	In-gel digestion → LC-MS/MS	Prefer Coomassie staining	Blank gel piece and/or neighboring band as control

Sample Requirements for Transcriptomics

Sample Class	Sample Type	Recommended	Minimum
Tissue Samples	Plant tissues (high nucleotide yield): young leaves, flowers, stems, anthers, young seeds	300 mg	20 mg
	Plant tissues (low nucleotide yield): roots, fruits, pericarp, mature seeds, high wax/lignin	300 mg	100 mg
	Human/Animal tissues (high yield): liver, spleen, thymus, heart, brain, lungs, kidneys	50 mg	10 mg
	Human/Animal tissues (low yield): skin, bone, fat	200 mg	100 mg
Cell Samples	Blood Cells	1 ml	0.5 ml
Cell Samples	Fresh cultured cells	5×10⁶ cells	1×10⁶ cells
Microorganisms	Bacteria	—	—
Microorganisms	Fungi	5×10⁶ cells or 0.3 g	1×10⁶ cells or 50 mg
RNA	Total RNA	Concentration ≥50 ng/μl; Total Amount ≥2 μg; Volume ≥15 μl; RIN ≥6

Sample Requirements for Microbiome

Sample Class	Sample Type	16S Sequencing		Metagenome Sequencing
Sample Class	Sample Type	Recommended	Minimum	Recommended	Minimum
Solid	Feces, Intestinal Contents	2 g	500 mg	200 mg	500 mg
	Swab	10 pieces	4 pieces	10 pieces	5 pieces
	Tissue	1 g	500 mg	/	/
	Water Body Filter Membrane (0.22–0.45 μm, ø 3–4 cm)	2 pieces	1 piece	3 pieces	1 piece
	Soil, Sludge, Sediment, Humus	3 g	0.5 g	500 mg	200 mg
Liquid	Rumen Fluid, Fermentation Broth, Tissue Fluid	3 ml, precipitate 1 g	1 ml, precipitate 0.5 g	5 ml, precipitate 300 mg	1 ml, precipitate 100 mg
	Saliva	5 ml	3 ml	5 ml	3 ml
	DNA	Conc. ≥5 ng/μl; Total ≥150 ng; Vol. ≥30 μl; OD 1.8–2.0		Conc. ≥10 ng/μl; Total ≥500 ng; Vol. ≥50 μl; OD 1.8–2.0

Sampling Guideline

Representative Sample	Sampling Method
Serum	Collect whole blood using serum separation tubes with clot activator. Allow to clot at room temperature for 60 minutes (do not shake). Centrifuge at 3000 rpm for 10 min at 4°C. Aliquot 200 μl supernatant into pre-labeled 2 ml tubes. Snap freeze in liquid nitrogen for 5–10 min, store at -80°C.
Plasma	Collect whole blood using anticoagulant tubes (EDTA, heparin, or citrate). Mix gently by inversion. Centrifuge at 3000 rpm for 10 min at 4°C. Aliquot 200 μl supernatant into pre-labeled 2 ml tubes. Snap freeze in liquid nitrogen for 5–10 min, store at -80°C.
Tissue	Rinse tissue briefly with PBS to remove blood/debris. Snap freeze in liquid nitrogen within 30 min of collection. Store at -80°C. Do not use RNAlater for metabolomics (it interferes with detection).
Urine	Collect mid-stream urine. Centrifuge at 4000 rpm for 10 min at 4°C to remove cells/debris. Aliquot supernatant and freeze at -80°C.
Cells	Wash cells 2–3 times with cold PBS. Snap freeze cell pellet in liquid nitrogen. Store at -80°C. Minimum 1×10⁶ cells recommended.
Feces	Collect fresh fecal samples. Freeze immediately in liquid nitrogen. Store at -80°C. Avoid freeze-thaw cycles. For microbiome, use sterile collection tubes.

Shipping Guideline

Dry Ice Booking: Book sufficient dry ice. Prepare at least 5 kg of dry ice for each transit day.
Sample Collection: Use appropriate tubes. Different services require separate tubes. Label clearly with markers that won't erase. Wrap tube cap with parafilm.
Sample Packaging: Wrap tubes in absorbent paper towels. Place in separate sealed plastic bags. Wrap each bag in bubble wrap.
Sample Information Form (SIF): Print and complete the SIF. Tube labels must match SIF labels. Place in sealed plastic bag inside the shipment.
Box Preparation: Place tubes and SIF in a polystyrene box with sufficient dry ice. Fill empty space with bubble wrap or packing material.
External Packaging: Place polystyrene box in a slightly larger cardboard box. Seal with packing tape. Affix a "Dry Ice" label to the outside.
Shipping Service: Use Next-Day delivery.
Notification: Email completed SIF, purchase order/quote, and tracking number to support-global@metwarebio.com.
Shipping Address:
Attn: Rui He
8A Henshaw Street, Woburn, MA 01801, USA
(781) 975-1541
Support-global@metwarebio.com

Frequently Asked Questions

For untargeted metabolomics, the recommended amount is 100 μl per sample, with a minimum of 20 μl. For targeted panels (e.g., steroid hormones, oxylipins), you may need 50–100 μl. Always provide extra volume if possible to allow for repeat analysis.

For DIA proteomics, the minimum sample amount depends on the tissue type. For normal animal tissues (liver, spleen, etc.), the minimum is 5 mg. For fatty tissue or skin, the minimum is 100 mg. For serum/plasma (without enrichment), as little as 1 μl is acceptable.

Wrap sample tubes in absorbent paper and sealed bags, then in bubble wrap. Place in a polystyrene box with at least 5 kg of dry ice per transit day. Put the polystyrene box inside a cardboard box, seal it, and attach a "Dry Ice" label. Use Next-Day delivery and email the tracking number to support-global@metwarebio.com.

A minimum of 3 biological replicates is required for all sample types. For animal studies, we recommend 3–6 replicates. For clinical samples, 6–10 replicates are recommended to account for individual variability.

If your sample type is not listed in our guidelines, please contact us. Our scientific team will evaluate your sample and provide specific guidance on collection, storage, and shipping requirements.

Can't Find Your Sample Type?

Our scientific team is here to help. Reach out with your specific requirements and we'll guide you through the process.