Isolated nuclei are widely used for a variety of experiments such as FACS , single nucleus analysis (such as RNA-seq and ATAC-seq), immunofluorescence staining, cell cycle and apoptosis research. Single cell RNA-seq is a powerful technology for studying complex cellular composition of tissues. However, Neurons are highly interconnected and it is very difficult to obtain single cells from neuronal tissues such as brain and spinal cord. It is even more difficult to isolate intact cells from frozen neuronal tissues. Due to these limitations, single cell RNA-seq is being substituted by single nucleus-seq. Traditional method for single nucleus isolation from neuronal tissue is relatively tedious and time consuming and the yield is usually low because it is difficult to get rid of contaminating myelin and other cellular components. This kit is specially designed to address the issues with a protocol that is simple, rapid and effective. Highly purified single nucleus can be obtained in about 30 min. In comparison to traditional method, the kit requires much less starting material and can handle a large range of sample size (1-25 mg). The buffers contain proprietary mix of detergents for efficient cell lysis. If the presence of detergent is not desirable, a detergent-free nuclei isolation kit is also available under cat# NI-024.
Image: Nuclei Isolated from zebrafish brain tissue Using BN-020 and stained with DAPI (False-coloured in green) (Courtesy Sema Elif Eski, Singh Lab, Université Libre de Bruxelles).
|Buffer A||15 ml|
|Buffer B||25 ml|
|Filter Cartridge with Collection Tubes||20 Units|
|Pestles for 1.5 ml Tubes||2 Units|
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