Minute™ Detergent-Free Single Nuclei Isolation Kit (20 Preps)

Manual & Protocol | Material Safety Data Sheets (MSDS)

*Invent Biotechnologies, Inc. is the inventor of the spin column-based cell fractionation technology (including single nuclei isolation). We have not authorized third parties to produce similar products using the technology. 

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Single-cell genomics requires high-quality and complete genetic materials from individual cells. However, intact single cells are hard to isolate from tissues, especially when frozen. In addition, dissociation-induced (either mechanical or enzymatic) population and expression bias are also problematic. It is more feasible to isolate intact nuclei to be used as the starting material for most single-cell genomics applications. 

The detergent-free nuclei isolation kit is designed to rapidly isolate intact nuclei from animal cultured cells or tissues (fresh or frozen). Intact nuclei can be isolated from the samples using proprietary spin-column-based technologies in less than 20 min without using tissue homogenizer and any detergents. The traditional method for nuclei isolation involves the use of non-ionic detergent, which has a tendency to cause unwanted nuclear aggregation. It is unclear why some nuclei are aggregated while others are not. It is also unknown whether non-aggregated nuclei are an unbiased representation of the whole nuclei population. Detergents also have the potential to damage the nuclear envelope resulting in leakage of nuclear matrix materials. For some cell/tissue types, nuclear envelop proteins could also be stripped off by the detergents. Minute^TM Detergent-Free Nuclei Isolation Kit provides a whole new way for nuclei isolation in contrast to traditional methods.

How it works: cells/tissues are first sensitized by buffer A before passing through the proprietary filter in a zigzag manner when high-speed centrifugal force is applied. The cells are ruptured when passing through the filter leaving intact native nuclei in the flow-through. The nuclei are separated from other small cell debris by low-speed centrifugation using the proprietary buffer B. The native and intact nuclei isolated can be used for a variety of downstream applications that include but not limited to: FACS analysis, single nucleus analysis (such as RNA-seq and ATAC-seq), immunofluorescence staining, cell cycle analysis, and/or apoptosis research.

For storage and transportation of isolated single nuclei, the following buffer is recommended:

WA-014: Minute™ Anti-Clumping Nuclei Storage Buffer 

NOTE: We also offer the following single nucleus isolation kits with a much cleaner background if the presence of detergent is not a concern:

SN-047: Minute™ Single Nucleus Isolation Kit for Tissues/Cells

BN-020: Minute™ Single Nucleus Isolation Kit for Neuronal Tissues/Cells

AN-029: Minute™ Nuclei and Cytosol Isolation Kit for Adipose Tissues/Cultured Adipocytes

Nuclei Isolated from zebrafish brain tissue Using NI-024 and stained with DAPI (False-coloured in green) (Courtesy Sema Elif Eski, Singh Lab, Université Libre de Bruxelles).

Kit includes:

1. Boerhan, R., Sun, W., Tian, N., Wang, Y., Lu, J., Li, C., ... & Zhou, Q. X. (2019). Fluorination on non-photolabile dppz ligand improving Ru (II) complex-based photoactivated chemotherapy. Dalton Transactions.

2. Eski, S.E., Dubois, C., Singh, S.P. NucleiIsolation from Whole Tissue usinga Detergent and Enzyme-FreeMethod. J. Vis. Exp. (160), e61471,doi:10.3791/61471 (2020).

3.  Yamada, S., Ko, T., Hatsuse, S., Nomura, S., Zhang, B., Dai, Z., ... & Komuro, I. (2022). Spatiotemporal transcriptome analysis reveals critical roles for mechano-sensing genes at the border zone in remodeling after myocardial infarction. Nature Cardiovascular Research, 1-12.

4.  Pernaa, N., Keskitalo, S., Chowdhury, I., Nissinen, A., Glumoff, V., Keski-Filppula, R., ... & Hautala, T. (2022). Heterozygous premature termination in zinc-finger domain of Krüppel-like factor 2 gene associates with dysregulated immunity. Frontiers in Immunology, 13, 6762.

5.  Rex, E. A., Seo, D., Chappidi, S., Pinkham, C., Brito, S., Embry, A., ... & Gammon, D. B. (2023). A FACT-ETS-1 Antiviral Response Pathway Restricts Viral Replication and is Countered by Poxvirus A51R Proteins. bioRxiv, 2023-02.

6. Yang, N., Occean, J. R., Melters, D. P., Shi, C., Wang, L., Stransky, S., ... & Sen, P. (2023). A hyper-quiescent chromatin state formed during aging is reversed by regeneration. Molecular Cell.

7.  Singh, A., Poling, H. M., Chaturvedi, P., Thorner, K., Sundaram, N., Kechele, D. O., ... & Helmrath, M. A. (2023). Transplanted human intestinal organoids: A resource for modeling human intestinal development. Development, dev-201416.

8. Rex, E. A., Seo, D., Chappidi, S., Pinkham, C., Brito Oliveira, S., Embry, A., ... & Gammon, D. B. (2024). FEAR antiviral response pathway is independent of interferons and countered by poxvirus proteins. Nature Microbiology, 1-19.

9.  Poling, H. M., Sundaram, N., Fisher, G. W., Singh, A., Shiley, J. R., Nattamai, K., ... & Helmrath, M. A. (2024). Human pluripotent stem cell-derived organoids repair damaged bowel in vivo. Cell Stem Cell.