The EXTRACTME DNA GEL-OUT KIT is designed for a rapid and efficient purification of DNA fragments directly from agarose gels (standard and low-melting point agarose gels run in either a TAE or TBE buffer). Agarose, ethidium bromide and other contaminants from a sample are effectively removed in the purification process. The kit enables the purification of DNA fragments from 50 bp to 30 kb, as well as plasmid and genomic DNA. However, purification of fragments smaller than 100 bp and larger than 10 kb will result in decreased recovery rates. The purified DNA can be used in common downstream applications. The purification protocol and buffer formulations were optimized for high yields and DNA purity. The product is intended for research use only.
DNA purification procedure utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first step, DNA fragment is excised from an agarose gel and incubated in GB Buffer, which enables gel fragment solubilization and protein degradation. As an added convenience, binding buffer contains a color indicator, which facilitates easy monitoring of the solution’s pH for optimal DNA binding. The two-step washing stage efficiently removes impurities and enzyme inhibitors. Purified DNA is eluted with the use either a low ionic strength buffer (Elution Buffer) or water (pH 7.0–9.0) and can be used directly in all downstream applications such as PCR, qPCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth or stored until ready to use.
Agarose fragment of up to 300 mg containing DNA
70–95%, depending on DNA fragment length (in the range of 100 bp – 10 kb)
DNA FRAGMENT LENGTH
100 bp – 10 kb
DNA fragments in the 50–100 bp and 10–30 kbp range can also be purified, as well as genomic and plasmid DNA. However, the efficiency will be decreased.
Approx. 25 μg DNA
A260/A280 ratio = 1.7 – 1.9
RECOMMENDATIONS AND IMPORTANT NOTES
An optimal volume of Elution Buffer used should be chosen in accordance with the amount of DNA in the sample and with final DNA concentration expected. The use of 30–100 μl of Elution Buffer is recommended. If high DNA concentration is desired elution’s volume may be reduced to 20 μl. It should be noted that this may reduce the efficiency of DNA retrieval. It is essential
to apply Elution Buffer precisely to the centre of the membrane. In order to maximize the DNA retrieval, heat Elution Buffer to 80°C and incubate it on the membrane for 10 minutes. If full DNA retrieval is required, elute DNA in 200 μl of Elution Buffer or perform a second elution. For the second elution, repeat steps 11–14 of the Isolation Protocol (section XI), placing the purification column in a new, sterile 1.5 ml Eppendorf tube. Elution Buffer does not contain EDTA, which may interfere with some enzymatic reactions.
GB Buffer contains an indicator, which enables pH monitoring. Yellow indicates that the solution’s pH is lower than 7.5, which guarantees optimal DNA binding with the membrane. When the pH is higher than 7.5, the solution turns pink. It usually happens, for example, when the running buffer for electrophoresis has been used several times or was incorrectly prepared. In such cases, it is essential to add 10 μl of 3 M sodium acetate (pH 5.2). It will lower the pH, enabling the solution to bind efficiently to the minicolumn membrane.
EXTRACTME DNA GEL-OUT KIT (EM08) - Discontinued - Manual
EXTRACTME DNA GEL-OUT KIT – TROUBLESHOOTING
Protect GB Buffer from the sunlight.
In order to avoid evaporation, ensure that the buffer bottles are tightly closed before storing.
The quality of each production batch (LOT) of the EXTRACTME DNA GEL-OUT KIT is tested with the use of standard QC procedures. Purified DNA concentration and quality are evaluated by gel electrophoresis and spectrophotometer.
EXTRACTME DNA GEL-OUT KIT (EM08) - Discontinued - MSDS
EXTRACTME DNA GEL-OUT KIT (EM08) - Discontinued - CoA L.695873
EXTRACTME DNA GEL-OUT KIT (EM08) - Discontinued - CoA L. 646573
EXTRACTME DNA GEL-OUT KIT (EM08) - Discontinued - CoA L. 706671
EXTRACTME DNA GEL-OUT KIT (EM08) - Discontinued - CoA L.656572
Investigation Of 16S rRNA Gene And Gene Segments For The Determination Of Probiotics
Authors Amar Cosic, Altijana Hromic Jahjefendic Products EXTRACTME GENOMIC DNA KIT (EM13), EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued Year 2019 Source Southeast Europe Journal of Soft Computing, 8, March 2019
New nuclease from extremely psychrophilic microorganism Psychromonas ingrahamii 37: identification and characterization
Authors Maciejewska, N., Walkusz, R., Olszewski, M., & Szymańska, A Products EXTRACTME DNA BACTERIA KIT (EM02), EXTRACTME DNA CLEAN-UP KIT (EM07) – Discontinued, EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued, PWO Polymerases, Plasmid DNA Isolation Kits Year 2018 Source Molecular biotechnology, 61(2), 122-133.
Liver Expression of Sulphotransferase 2A1 Enzyme Is Impaired in Patients with Primary Sclerosing Cholangitis: Lack of the Response to Enhanced Expression of PXR
Authors Ewa Wunsch, Marta Klak, Urszula Wasik, Malgorzata Milkiewicz, Malgorzata Blatkiewicz, Elzbieta Urasinska, Olivier Barbier, Dariusz Bielicki, Dimitrios P. Bogdanos, Elwyn Elias and Piotr Milkiewicz Products EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued Year 2015 Source Hindawi Publishing Corporation Journal of Immunology Research Volume 2015, Article ID 571353, 8 pages
Mutation of the PIK3CA gene as a prognostic factor in patients with colorectal cancer
Authors Stec Rafał, Semeniuk-Wojtaś Aleksandra, Charkiewicz Radosław, Bodnar Lubomir, Korniluk Jan, Smoter Marta, Chyczewski Lech, Nikliński Jacek, Szczylik Cezary. Products EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued Year 2015 Source ONCOLOGY LETTERS 10: 1423-1429, 2015
Identification and characterization of single-stranded DNA-binding protein from the facultative psychrophilic bacteria Pseudoalteromonas haloplanktis
Authors Marcin Olszewski, Marta Nowak, Anna Cyranka-Czaja, Józef Kur Products EXTRACTME DNA BACTERIA KIT (EM02), EXTRACTME DNA CLEAN-UP KIT (EM07) – Discontinued, EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued, PWO Polymerases, PCR & Real-Time PCR Year 2014 Source Microbiological Research 169 (2014) 139– 147
Characterization of single-stranded DNA-binding proteins from the psychrophilic bacteria Desulfotalea psychrophila, Flavobacterium psychrophilum, Psychrobacter arcticus, Psychrobacter cryohalolentis, Psychromonas ingrahamii, Psychroflexus torquis, and Photobacterium profundum
Authors Marta Nowak, Marcin Olszewski, Marta Śpibida and Józef Kur Products EXTRACTME DNA BACTERIA KIT (EM02), EXTRACTME DNA CLEAN-UP KIT (EM07) – Discontinued, EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued, PCR & Real-Time PCR, PWO Polymerases Year 2014 Source Nowak et al. BMC Microbiology 2014, 14:91