User Manual

BioMasher is a disposable homogenizer.
BioMasher is used to disrupt the tissue sample when extracting RNA, DNA, and proteinsfrom the sample.
The BioMasher lineup includes the BioMasher I, II, III, V, and SP models.
BioMasher I has five variations: with or without an O-ring and by filter tube pore size.

BioMasher I
BioMasher I consists of a filter tube and a pestle. The
sample is placed in the filter tube and the pestle isinserted from above. Centrifugal force is applied with acentrifuge allowing the pestle to crush the sample withthis force. This allows the sample to pass through thefilter of the filter tube, enabling the sample to bedisrupted. The fibrous membrane component of thesample will be caught by the filter.
The pestle has a cross-shaped blade at the tip. By
pushing and turning the pestle manually, the sample,
which is hard to disrupt by centrifugation alone, can beground.
<Variation>★ With an O-ring
BioMasher I with an O-ring is suitable forhomogenizing soft tissue with centrifugal force alone.
In particular, it is suitable for simultaneous processingof many samples and for infectious samples.
★ Without an O-ringHard tissue may not be homogenized sufficiently bycentrifugal force alone. The model without an O-ring issuitable when grinding the sample on the filter area
before centrifugation to enhance disruption efficiency.
＊The material of the filter is available made of either
polyethylene (PE) or polypropylene (PP).
The PE filter has smaller pore size than the PP filter
resulting in finer disruption with the PE filter.
* DO NOT add the buffer in the filter tube before
centrifuge. Otherwise the filter may be clogged, and thetube may be damaged. For only samples with lots offibrous components with little water content, such as
epidermis, mesentery, tail etc., add a small amount ofbuffer to extract and filtrate the target molecules.

<protocol>-1-

BioMasher II
BioMasher II consists of a 1.5 mL microtube and apestle. The sample and extraction buffer are placed in
the tube, and ground to prepare the homogenate.
The microtube and pestle are molded so that they fit
without any space between them. The texture of theinner wall of the tube and the outer periphery of the tipof the pestle enhances the efficiency of the disruptionof the sample.
The pestle has a splash guard, which is designed to fit
the tube without unnecessary spaces.
BioMasher II can be used to disrupt various samples,such as animal tissue/organ, plants (stems, roots,seeds), and insects.
★For samples that are difficult to disrupt, an exclusiveelectric stirrer (Nippi PowerMasher II) is available foreasy disruption.
★DO NOT autoclave BioMasher II. In the casesterilization is required, please use a sterilized product.

<protocol>-2-BioMasher SP
BioMasher SP is a larger volume version of BioMasher
II. This model has enhanced disruption efficacy fromthe combination of the 15 mL tube with an abrasive
inner wall and a stirring pestle with a textured tip.
The shovel-like shape cutout and the helical groove of
the pestle effectively trap and homogenize the tissue.
BioMasher SP is suitable for disruption of animal
tissue/organ, plants (stems, roots, seeds), and insects.
★For samples difficult to disrupt, an exclusive electricstirrer (Nippi PowerMasher II) is available for easy
disruption.
★ DO NOT autoclave BioMasher II. In the casesterilization is required, please use a sterilized product.

<protocol>-3-BioMasher III
BioMasher III has the features of both BioMasher I andII. This model consists of a filter tube with an abrasive
surface inner wall and a pestle with a textured surface.
After grinding the sample in the filter tube using thepestle, centrifuge the extraction liquid.
The extracted sample will be filtered and caught in thetube.
Since the fibrous membrane component get trappedby the filter, BioMasher III is recommended forhomogenization of highly fibrous samples.
★For samples that are difficult to homogenize, anexclusive electric stirrer (Nippi PowerMasher II) can beused for ease.

<protocol>-4-BioMasher V
BioMasher V is a larger volume version of BioMasher
III.
After homogenizing the sample with a combination ofthe tube with a textured inner wall and the stirring
pestle with an abrasive surface, centrifuge and filtratewith the filter.
Recommended for homogenizing highly fibroussamples.
★For samples difficult to homogenize, an exclusiveelectric stirrer (PowerMasher II) is available for ease.

<protocol>-5-PowerMasher II helps the disruption of large amounts of samples and disruption that isdifficult to complete manually. The device is attached to the stirring pestle of the BioMasher
II, III, V, and SP to allow electrical stirring.
Geared high torque type with a rotation speed of 450 rpm.
*Two AA dry-cell batteries are required.

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BioMasher Series Model Suitability by Tissue Type (1)Soft Tissue

Brain, medulla oblongata, kidneys, liver, spleen, etc.

Containment is required.
(e.g. infectious substance)

YESBM I with O-ring→ P 9-(1)*Some agent-treatedsamples may be difficult
to disrupt.

NOSimultaneous
processing of many
samples is required.

NONO

highly fibrous sample

YES

YES

BM II or BMSP
→ P 10If disruption volume
is small: BM II
If disruption volumeis large: BMSPBM III or BM V
→ P 11
If disruption volume is small: BM III
If disruption volume is large: BM V

YES

More efficient
extraction is required.

BM I without O-ring→NOBM I with O-ring→P 9-(1)-7-P 9-(2)BioMasher Series Model Suitability by Tissue Type (2)Hard Tissue

Epithelial tissue, digestive organs, heart, muscle, cartilage, etc.

Containment is required.
(e.g. infectious substance)

YES
BM I with O-ring→P 9-(1)NO

YES

More efficiency isrequired.

Highly fibrous sample

NOYESNOBM II or BMSP
→ P 10If disruption volume
is small: BM II
If disruption volume
is large: BMSPBM I without O-ring

BM III or BM V→→ P 11
If disruption volume is small: BM III
If disruption volume is large: BM V

P 9-(2)-8-

BioMasher I Protocol(1) For the liver, kidneys, cerebrum, cerebellum, and spleen (fresh sample)→Use a pestle with an O-ring.
1. Set a filter tube into the recovery tube and put 50–100 mg of the tissue to disrupt in
the filter tube.
2. Insert the pestle with the O-ring into the filter tube and push it to the end.
3. Centrifuge at 15,000 x g for 30 seconds.
4. Discard the filter tube and pestle.
5. Add 1 mL of disruption buffer in the recovery tube and vortex.
(2) For the small intestine, large intestine, lung, tail, muscle, seminal vesicle,
gallbladder, salivary glands, preputial gland, heart, blood vessels, and agent-treatedorgans→ Use a pestle without an O-ring.
1. Set a filter tube into the recovery tube and put 50–100 mg of the tissue to disrupt in
the filter tube.
2. Add 100 μL of disruption buffer in the filter tube and insert the pestle without an Oring.
3. Disrupt the tissue by rotating the pestle while pushing it into the filter area of thefilter tube.
4. Centrifuge at 15,000 x g for 30 seconds.
5. Discard the pestle and add 900 μL of disruption buffer to the filter tube.
6. Centrifuge at 15,000 g for 30 seconds.
7. Discard the filter tube.
8. Close the cap of the recovery tube and vortex.
References which BioMasher I is used.
1) Yamamoto, T. et al. (2012) Improved RNA extraction method using the BioMasher andBioMasher power-plus. Journal of Veterinary Medical Science, 74: 1561-7
2) Yamamoto, T. et al. (2013) Sensitivity and specificity of a commercial BSE kit for thedetection of ovine scrapie. Animal Science Journal, 84: 508-123) Aboelenain, M. et al. (2015) Status of autophagy, lysosome activity and apoptosis during
corpus luteum regression in cattle. Journal of Reproduction and Development, 61: 22936
4) Lee, B.S. et al. (2017) Hypothermia decreased the expression of heat shock proteins in
neonatal rat model of hypoxic ischemic encephalopathy. Cell Stress and Chaperones,
22: 409–155) Ikeda, R. et al. (2015) Knockdown of aquaporin-8 induces mitochondrial dysfunction in3T3-L1 cells. Biochemistry and Biophysics Reports, 4: 187-956) Uchida, T. et al. (2017) Abnormal γ-aminobutyric acid neurotransmission in a Kcnq2
model of early onset epilepsy. Epilesia, 58: 1430-397) Nakayama, T. et al. (2017) Detection of fungi from an indoor environment using loopmediated isothermal amplification (LAMP) Method. Biocontrol Science, 22: 97-1048) Roberts, B.R. et al. (2017) Biochemically-defined pools of amyloid-β in sporadic
Alzheimer’s disease: correlation with amyloid PET. Brain, 140: 1486–98-9-BioMasher II Protocol
Samples to disrupt: Animal tissues/organs, plants (stems, roots, seeds), insects,etc.
1. Put 50–100 mg of tissue to disrupt in the tube included in BioMasher II.
2. Add 250 μL of disruption buffer.
3. Insert the pestle and disrupt the tissue while pressing the pestle to the side ofthe tube.
4. Discard the pestle and add 750 μL of disruption buffer.
5. Vortex.
BioMasher SP Protocol
Samples to disrupt: Animal tissues/organs, plants (stems, roots, seeds), insects,etc.
1. Put the tissue to disrupt in the tube included in BioMasher SP.
2. Add disruption buffer.
3. Insert the pestle and disrupt the tissue while pressing the pestle to the side ofthe tube.
4. Discard the pestle and add disruption buffer.
5. Vortex.

References which BioMasher II or SP is used.
1) Jinnai, M. et al. (2017) Production of a novel monoclonal antibody applicable for an
immunochromatographic assay for Kudoa septempunctata spores contaminating the raw olive
flounder (Paralichthys olivaceus). International Journal of Food Microbiology, 259: 59-67
2) Jasper Elvin James et al. (2016) In vitro antifungal susceptibility of neoscytalidium dimidiatum
cinical isolates from malaysia. Mycopathologia, 182: 305–133) Gilbert, J. R., et al. (2017) Resequencing of the Col1A1 gene of Oryctolagus cuniculus
identifies splicing errors and single nucleotide polymorphisms. Genes & Genomics, 39: 549–554) Kobayashi, D. et al. (2017) Detection of a novel putative phlebovirus and first isolation ofDugbe virus from ticks in Accra, Ghana. Ticks and Tick-borne Diseases, 8: 640-455) Kwon, D.H. et al. (2017) Genetic diversity and structure in apple-infesting pests of Carposina
sasakii, Grapholita dimorpha and Grapholita molesta in Korea. Journal of Asia-Pacific
Entomology, 20: 13-166) Isobe, K. et al. (2017) Platelet-rich fibrin prepared from stored whole-blood samples.
International Journal of Implant Dentistry, 2017: 3-67) Ishizawa, H. et al. (2017) Differential oxidative and antioxidative response of duckweed
Lemna minor toward plant growth promoting/inhibiting bacteria. Plant Physiology andBiochemistry, 118: 667-738) Matsubara, T. et al. (2017) Regulation of osteoclast differentiation and actin ring formation by
the cytolinker protein plectin. Biochemical and Biophysical Research Communications, 489:
472–76-10-

BioMasher III Protocol
Samples to disrupt: Among animal tissues/organs, plants (stems, roots, seeds),
insects, etc., samples with much fiber
1. 1.Put the sample in BioMasher III and disrupt it with 100 μL of disruption
buffer.
2. 2.Wash out the tissue section attached to the pestle with 200 μL ofdisruption buffer in the filter tube.
3. Discard the pestle and centrifuge at 10,000 x g for 30 seconds at room
temperature.
4. Add 700 μL of disruption buffer.
5. Vortex.
BioMasher V Protocol
Samples to disrupt: Among animal tissues/organs, plants (stems, roots, seeds),
insects etc., samples with much fiber
1. Put the sample in BioMasher V and disrupt it with disruption buffer.
2. Wash out the tissue section attached to the pestle with disruption buffer inthe filter tube.
3. Discard the pestle and centrifuge at 6,000 g for 30 seconds at room
temperature.
4. Add disruption buffer.
5. Vortex.

References which BioMasher III or V is used.
1) Alqaydi, M. et al. (2016) Quantitative and qualitative study of STR DNA from ethanol andformalin fixed tissues. Forensic Science International, 262: 18-292) Yoshizawa, T. et al. (2016) Effects of adrenomedullin on doxorubicin-induced cardiac damage
in mice. Biological and Pharmaceutical Bulletin, 39: 737-463) Kimoto, M. et al. (2016) DNA aptamer generation by genetic alphabet expansion SELEX(ExSELEX) using an unnatural base pair system. Nucleic Acid Aptamers, Part of the Methods
in Molecular Biology book series, MIMB, 1380: 47-60-11-FAQ
Q1. In terms of sample volume, which model of the BioMasher isappropriate?
→ Our recommendation is as below.
Sample size
BM I: 50 mg
BM II: 100 mg
BM III: 50 mg
BM V: 500 mg
BM SP: 1,000 mg
Q2. What is the centrifugation resistance of the BioMasher?
→ Although it depends on sample, Please try the following conditions asa standard.
BM I: 10,000-15,000 g, 10-30 seconds
BM II: 10,000-15,000 g, 10-30 seconds
BM III: 6,000-10,000 g, 10-30 seconds
BM V: 3,000-6,000 g, 5 minutes
BM SP: 3,000-6,000 g, 5 minutes
Q3. What is the lowest temperature BioMasher can bear?
→ The material of the tube is PP, and its freezing resistance is approximately-60°C to -40°C.
Cooling of the sample with liquid nitrogen together with the tube may cause
damage.

Q4. Contact for other inquiries→ Please access the following URL, which has the link to the inquiry
form.
https://www.nippi-inc.co.jp/tabid/143/Default.aspx-12-