Sample Submission Guideline for NGS Grade Nucleic Acid Extraction: NGS-1000 series
Sample Type/Origin

NGS Grade DNA extraction

Animal Tissue

Option 1: Tissue• Excise target tissue from animal• Rinse tissue with 1X PBS to remove all traces of blood• Weight: 50 - 100 mg of sample**Cut and weighed sample must be completed within 3 minutes after tissue removal from animal to ensure the tissue is preserved without degradation.
• Snap freeze sample in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

NGS Grade RNA extraction

Option 2: Tissue in fine powder• Excise target tissue from animal• Rinse tissue with 1X PBS to remove all traces of blood• Quickly grind tissue into fine powder with liquid nitrogen• Weight: 50 - 100 mg of sample• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice
Option 3: Tissue in DNA/RNA tissue stabilization solution• Excise target tissue from animal• Cut into slices less than 0.5 cm thickness.
• Put fresh tissue into screw cap tube (1.5/2.0/15/50 ml).
• Weight: 50 - 100 mg of sample**Cut and weighed sample must be completed within 3 minutes after tissue removal from animal to ensure the tissue is preserved without degradation.
• Preserve the sample according to the guidelines of the stabilization reagent used, following the ratio of stabilization reagent volume to sample size to ensure effective sample preservation.
• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Cultured Cells

Option 1: Cell pellet• Cells up to 1 x 107 (1 to 2 quantity of T25 flask, at 70% confluency)• Collect pellet from culture, wash cell pellet with PBS and remove PBS completely• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice
Option 2: Cell pellet in DNA/RNA stabilization solution
7• Cells up to 1 x 10 (1 to 2 quantity of T25 flask, at 70% confluency)• Collect pellet from culture, wash cell pellet with PBS and remove PBS completely• Preserve the sample according to the guidelines of the stabilization reagent used, following the ratio of stabilization reagent volume to sample size to ensure effective sample preservation.
• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Animal Tissue/ Cultured Cells for
Mitochondria DNA (mtDNA) Extraction

Animal Tissue• Excise target tissue from animal• Rinse tissue with 1X PBS to remove all traces of blood• Weight: 50 - 100 mg of sample• Snap freeze sample in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Not applicable.

Cell pellet
7• Cells up to 1 x 10 (1 to 2 quantity of T25 flask, at 70% confluency)• Collect pellet from culture, wash cell pellet with PBS and remove PBS completely• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice
Bacteria

Bacterial pellet• Collect pellet from 10 - 30 ml overnight culture, remove culture medium completely• Weight: 200 - 500 mg of sample• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Bacterial pellet• Dilute overnight culture to 1:50 ratio with culture media, continue grow for 3 - 5 hours**Do not directly use overnight culture. Reduce the dilution factor if slow-growing bacterial is used.
• Collect the pellet from culture, remove culture medium completely• Weight: 200 - 500 mg of sample• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry iceYeast

Yeast pellet• Collect pellet from culture, remove culture medium completely• Weight: 200 - 500 mg of sample• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Yeast pellet• Dilute overnight culture to 1:50 ratio with culture media, continue grow for 4 - 5 hours**Do not directly use overnight culture.
• Collect the pellet from culture, remove culture medium completely• Weight: 200 - 500 mg of sample• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Sample Submission Guideline for NGS Grade Nucleic Acid Extraction: NGS-1000 series
Sample Type/Origin

NGS Grade DNA extraction

Fungi

Fungal pellet• Collect pellet from culture, remove culture medium completely• Weight: 0.5 - 1 g of sample• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3 - 5• Store sample in -80 °C freezer• Shipping condition: Dry ice

NGS Grade RNA extraction

Algae

Microalgae pellet• Collect pellet from culture, remove culture medium completely• Weight: 0.5 - 1 g of sample• Snap freeze pellet in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry iceBloodA) Genomic DNA extraction from blood

Total RNA extraction from blood

Option 1: Whole Blood, *Malaysia Customer only
• Volume: 3 ml freshly collected blood in EDTA Tube
• Blood samples to be submitted immediately after withdrawal.
• Results will not be guaranteed for blood samples that are more than 3 days old• No. of replicates: 3• Shipping condition:i) 4°C or Blue ice (reach Apical Scientific within 3 days)ii) Dry ice (reach Apical Scientific > 3 days)

Option 1: Whole Blood in Tempus™ Blood RNA Tube
• Collect up to 3 ml blood in Tempus™ Blood RNA Tube
• Shake vigorously for 10 seconds to mix sample with stabilizing reagent in the Tempus Tube
• No. of replicates: 2• Store sample in 4 °C fridge (up to 7 days) or -20°C/-80°C freezer (> 7 days).
• Shipping condition:i) 4°C or Blue ice (reach Apical Scientific within 5 days)ii) Dry ice (reach Apical Scientific > 5 days), avoid direct contact of sample with dry ice!!

Option 2: Buffy Coat
• Collect up to 3 ml blood in EDTA tube, centrifuge the blood with 2,500 x g for 10 minutes using swing bucket
centrifuge• Remove the plasma with 1 ml syringe, 21G needle• Collect the buffy coat in 1.5/2 ml tube
• No. of replicates: 3• Store buffy coat in -80 °C freezer• Shipping condition: Dry ice

Option 2: White Blood Cell pellet• Collect up to 3 ml blood in EDTA tube
• Must preprocess the blood sample within the same day of sample collection.
• Pretreat the whole blood sample with RBC lysis buffer (1st BASE, K.BUF-9101-100ml), follow Appendix Protocol below
for pretreatment.
• No. of replicates: 3• Store the treated dry WBC pellet in -80 °C freezer. Alternatively, the WBC pellet can be stored in RNAlater solution.
• Shipping condition: Dry ice

Option 3: White Blood Cell pellet• Volume: up to 3 ml freshly collected blood in EDTA Tube
• Pretreat the whole blood sample with RBC lysis buffer (1st BASE, K.BUF-9101-100ml), follow Appendix Protocol belowfor pretreatment.
• No. of replicates: 3• Store the treated dry WBC pellet in -80 °C freezer. Alternatively, the WBC pellet can be stored in RNAlater solution.
• Shipping condition: Dry ice

InsectAppendix Protocol: Pretreatment of whole blood sample with RBC lysis buffer (1st BASE, K.BUF-9101-100ml)

Appendix Protocol: Pretreatment of whole blood sample with RBC lysis buffer (1st BASE, K.BUF-9101-100ml)

B) For cell-free DNA (cfDNA) extraction from blood plasma, please contact us to enquire.

Not applicable.

Option 1: Insect• Immerse the live insect in absolute ethanol until the insect stops moving• Blot dry the insect• Weight: 50 - 100 mg of sample• Snap freeze insect in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice
Option 2: Insect in fine powder• Immerse the live insect in absolute ethanol until the insect stops moving• Blot dry the insect• Quickly grind the insect into fine powder with liquid nitrogen• Weight: 50 - 100 mg of sample• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice
Option 3: Insect in DNA/RNA tissue stabilization solution• Immerse the live insect in absolute ethanol until the insect stops moving• Blot dry the insect• Put fresh insect into screw cap tube (1.5/2.0/15/50 ml).
• Weight: 50 - 100 mg of sample• Preserve the sample according to the guidelines of the stabilization reagent used, following the ratio of stabilization reagent volume to sample size to ensure effective sample preservation.
• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Sample Submission Guideline for NGS Grade Nucleic Acid Extraction: NGS-1000 series
Sample Type/Origin

NGS Grade DNA extraction

Plant

Fresh leaf/needle/stem/root, prefer young leaf/needle• Rinse with clean water to remove dirt attached to the plant sample• Cut sample into small pieces (< 0.5 cm), keep in 50 ml centrifuge tube
• Weight:i) 2 - 5 g wet weight ORii) 2 - 5 g liquid nitrogen ground fine powder**Cut and weighed sample must be completed within 3 minutes after tissue removal from plant to ensure the tissue is preserved without degradation.
• Snap freeze in liquid nitrogen bath for 10 minutes• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice<!> Avoid pooling different plant parts/organs into the same tube, as lysis procedure is different for each part.

Plant for Chloroplast DNA (cpDNA)
Extraction

Fresh green leaves without brown spots and holes• Weight: Harvest ≥ 30 g of fresh green leaves• Wrap leaves with aluminium foil
• No. of replicates: 3• Storage/Shipping condition: 4°C or Blue ice in dark condition<!> The sample must reach Apical Scientific within 48 - 72 hours upon sample collection.
Suggest to prearrange the best day for sample submission with our Customer Care team.

Not applicable.

Soil/Sludge

For Amplicon Sequencing application• Weight: 0.5 - 1 g of sample in screw cap tube
• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice• Collect soil sample in soil preservation solution, e.g. Qiagen LifeGuard Soil Preservation solution.
• Weigh 2 g of soil sample in 15 ml screw cap tube
• Add 5 ml of soil preservation solution• Vortex or invert tube by hand until the entire soil sample and preservation solution are mixed well.
Excess preservation solution should be sitting on top of the soil sample.
• No. of replicates: 3• Store sample in 4°C for overnight, transfer to -20°C freezer the next day.
• Shipping condition: Dry ice*Depends on soil types, additional sample replicates of 4 to 8 may be required in order to get sufficient RNA amountfor downstream applications.

For Shotgun Metagenomics application, we recommend to collect soil sample in soil preservation solution, e.g.
Qiagen LifeGuard Soil Preservation solution.
• Collect soil sample in soil preservation solution, e.g. Qiagen LifeGuard Soil Preservation solution.
• Weigh 2 g of soil sample in 15 ml screw cap tube
• Add 5 ml of soil preservation solution• Vortex or invert tube by hand until the entire soil sample and preservation solution are mixed well.
Excess preservation solution should be sitting on top of the soil sample.
• No. of replicates: 3• Store sample in 4°C for overnight, transfer to -20°C freezer the next day.
• Shipping condition: Dry ice*Depends on soil types, additional sample replicates of 4 to 8 may be required in order to get sufficient DNA amountfor downstream applications.

NGS Grade RNA extraction

Stool• Weight: 200 - 500 mg of sample in screw cap tube
• Label sample with printed labelling• No. of replicates: 3• Store sample in -80 °C freezer• Shipping condition: Dry ice

Manure• Weight: 200 - 500 mg of sample in screw cap tube
• Label sample with printed labelling• Store sample in -80 °C freezer• No. of replicates: 3• Shipping condition: Dry iceNot available.

WaterOption 1: Filtered content on 0.22 µm membrane filter• Prefilter water sample with larger pore size cell strainer (e.g. 100 µm) to remove large size debris• Continue vacuum filtration of 300 - 500 ml* water sample with 0.22 µm pore size, 25/47 mm membrane filter*If the sample has low turbidity (contained less filtered content), please continue to filter to reach volume of 1.5 L intotal
• Use a sterile scalpel blade to remove the membrane filter• Insert the membrane filter into a new tube by rolling the membrane using sterile forceps, with the side containing
trapped sample facing inward of the tube
• No. of replicates: 3• Store membrane filter in -80 °C freezer• Shipping condition: Dry ice

Not available.

Option 2: Water sample• Volume: 500 ml****If the sample has low turbidity, please submit at least triplicates, i.e. 500 ml x 3 = 1.5 L• Store sample in dark in 4°C in a screw cap bottle• Shipping condition: 4°C or Blue ice

Sample Submission Guideline for NGS Grade Nucleic Acid Extraction: NGS-1000 series
Sample Type/OriginNGS Grade DNA extraction

NGS Grade RNA extraction

FoodFood (Solid)• Weight: 10 g sample in screw cap tube/container/zip lock bag• No. of replicates: 2• Shipping condition: follow the food storage condition, as long as the food is not rotten during shipmenta) If it was stored in 4°C, send in blue iceb) If it was stored in freezer, send in dry ice

Not available.

Food (Liquid)• Weight: 20 - 50 ml sample in screw cap tube
• No. of replicates: 2• Shipping condition: follow the food storage condition, as long as the food is not rotten during shipmenta) If it was stored in 4°C, send in blue iceb) If it was stored in freezer, send in dry ice
Swab sample(Buccal swab/ enviromental swab)

Option 1:• Collect sample using commercial swab sample collection kit• Follow collection kit guidelines to preserve the swab sample• No. of replicates: 3• Storage condition: Follow the guidelines from the commercial kit• Shipping condition: Follow the guidelines from the commercial kit• Collect sample using commercial swab sample collection kit (for RNA application)• Follow collection kit guidelines to preserve the swab sample• No. of replicates: 3• Storage condition: Follow the guidelines from the commercial kit• Shipping condition: Follow the guidelines from the commercial kit

Option 2:• Collect sample using sterile swab
• (Optional) Store swab in 1X PBS• No. of replicates: 3• Storage condition: -80°C freezer• Shipping condition: Dry ice
SalivaOption 1:
Option 1:• Suggest to follow commercially available saliva collection kit guidelines for sample collection, preservation, storage and • Suggest to follow commercially available saliva collection kit guidelines for sample collection, preservation, storage andshipping.
shipping.
For example, DNA Genotek, OMNIgene,SALIVA DNA and RNA device.
For example, DNA Genotek, OMNIgene, SALIVA DNA and RNA device.
• No. of replicates: 2• No. of replicates: 2• Storage/Shipping condition: Follow kit recommendation• Storage/Shipping condition: Follow kit recommendation
Option 2:• Collect 2 ml saliva in a new 50 ml centrifuge tube
• Add 10 ml 1X PBS.
• Shake vigorously to mix for at least 20 seconds.
• Immediately centrifuge at 2,000 x g for 5 minutes at room temperature to pellet cells.
• Decant the supernatant without delay.
• No. of replicates: 2• Store dry pellet in -80 °C freezer• Shipping condition: Dry iceOthers

Please enquire.

How to snap freeze sample?
• Place the sample into a sterile empty tube
(1.5/2 ml tube with safety lock cap or 15/50 ml screw cap centrifuge tube)• Close the tube
• Immediately submerge the tube into liquid nitrogen bath
• The sample should be froze for at least 10 minutes• Store sample in -80 °C freezer

Option 2:• Collect 2 ml fresh saliva in a 50 ml centrifuge tube
• No. of replicates: 2• Add 2 ml RNAlater solution.
• Vortex vigorously to mix thoroughly.
• Incubate sample at 4°C fridge for overnight.
• Store in -80°C freezer the next day.
• Shipping condition: Dry ice