{"product_id":"nick-columns-17085501","title":"NICK Columns","description":"\u003cp\u003eNICK Columns are used for the purification of nick-translated DNA fragments and for separation of any labeled probe from unincorporated labeled nucleotides.\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003eFor rapid purification of labeled DNA (≥ 20 bases in length) from unincorporated radiolabeled nucleotides using gravity-flow chromatography\u003c\/li\u003e\n\u003cli\u003ePrepacked with Sephadex G-50 DNA Grade (see Sephadex G-50 DNA Grade and Sephadex G-100 DNA Grade) in distilled water with 0.15% Kathon CG\/ICP Biocide\u003c\/li\u003e\n\u003cli\u003eCan accommodate sample volumes up to 100 μl\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eNICK columns are ready for immediate use and operate by gravity flow. Tests show that at least 97% of radioactivity applied in a nick-translated mixture elutes in two well-separated peaks, corresponding to labeled DNA molecules and unincorporated nucleotides, respectively. At least 90% of the applied DNA is recovered and can then be used in DNA hybridization protocols. Maximum sample volume is 100 μL.\u003c\/p\u003e\n\n\u003cp\u003eNick translation is a technique in which DNA Polymerase I is used to replace some of the nucleotides of a DNA sequence with their labeled analogues, creating a tagged DNA sequence which can be used as a probe in fluorescent in situ hybridization (FISH) or blotting techniques. It can also be used for radioactive labeling in techniques such as Southern blotting.\u003c\/p\u003e\n\n\u003cp\u003eThis process is called nick translation because the DNA to be labelled is treated with DNase to produce single-stranded \"nicks.\" This is followed by replacement in nicked sites by DNA polymerase I, which elongates the 3' end, removing nucleotides by 5'-3' exonuclease activity, and replacing them with dNTPs. To label a DNA fragment for use as a probe in blotting one of the incorporated nucleotides provided in the reaction is radiolabeled, or a fluorophore can be attached instead for fluorescent labelling, or is used as an antigen for immunodetection.\u003c\/p\u003e\n\n\u003cp\u003eSephadex G-50 DNA Grade can be used in a wide range of applications, including desalting DNA, buffer exchange, and removal of unincorporated nucleotides from end-labelled oligonucleotides. Sephadex G-50 can be packed into empty MicroSpin columns for use in these applications.\u003c\/p\u003e\n\n\u003cp\u003eSephadex G-50 is one of five different G-types ranging from G-10 for small molecules to G-75 for larger molecules. Sephadex G-50 is a well-established gel filtration resin for desalting and buffer exchange of biomolecules \u0026gt; 30 000 molecular weight, and with a spin protocol can be used for DNA and oligo purification of molecules greater than 20 bases in length. Alternatively, Sephadex G-25 can be used. Sephadex G-25 has an exclusion limit of approximately Mr 5000, and with a spin protocol this means it can be used for any DNA greater than 10 bases in length.\u003c\/p\u003e\n\n\u003cp\u003eBoth types of Sephadex are therefore highly suitable for the purification of oligonucleotides or very small DNA fragments following synthesis or in radioactive labeling.\u003c\/p\u003e\n\n\u003cp\u003e\u003ca href=\"\/shop\/molecular-biology\/purification\/gel-filtration-columns\/illustra-sephadex-g-25-dna-grade-p-00054\"\u003eSephadex G-25 DNA Grade\u003c\/a\u003e and \u003ca href=\"\/shop\/molecular-biology\/purification\/gel-filtration-columns\/illustra-sephadex-g-50-dna-grade-p-00113\"\u003eSephadex G-50 DNA Grade\u003c\/a\u003e are sold separately.\u003c\/p\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cul\u003e\u003cli\u003e\u003ca href=\"https:\/\/d3.cytivalifesciences.com\/prod\/IFU\/17085501AC.pdf\"\u003eInstructions for use (PDF)\u003c\/a\u003e\u003c\/li\u003e\u003c\/ul\u003e","brand":"Cytiva","offers":[{"title":"20 columns","offer_id":45982269931599,"sku":"17085501","price":1377.0,"currency_code":"ILS","in_stock":true},{"title":"50 columns","offer_id":45982269964367,"sku":"17085502","price":2975.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/digi-30177-original-png_626fc002-9cca-4e03-81f2-925904968ec1.png?v=1785959655","url":"https:\/\/www.danyel.co.il\/products\/nick-columns-17085501","provider":"danyel biotech","version":"1.0","type":"link"}