32,00 € – 509,00 €The EXTRACTME GENOMIC DNA KIT is designed for a rapid and efficient purification of high quality genomic, mitochondrial, bacterial, parasite or viral DNA from solid tissues, physiological fluids, fresh and frozen blood, human and animal mucosa membrane swabs, semen, hair, rodent tails, insects, bacteria, yeast and cell cultures. The isolation protocol and buffer formulations have been optimized for high isolation efficiency and DNA purity. The product is intended for research use only.
DNA purification procedure consists of four steps and utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first step, biological material is lysed by Proteinase K in optimized GL Buffer. At this stage all the cellular membranes and proteins are degraded. When it is necessary to remove RNA the use of RNase A is recommended. After the addition of chaotropic salts, the lysate is applied to the purification minicolumns membrane and the DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. Purified DNA is eluted with the use of either a low ionic strength buffer (Elution Buffer) or water (pH 7.0–9.0) and may be used directly in all downstream applications such as PCR, quantitive real-time PCR, pharmacogenic research, Southern blotting, single-nucleotide polymorphism (SNP), short tandem repeat (STR) genotyping, DNA sequencing, enzymatic restriction, ligation and so forth, or stored until ready to use.
- fresh or frozen solid tissue: 1–30 mg
- formalin-preserved tissue: 1–30 mg
- paraffin-embedded tissue: 1–30 mg
- physiological fluids (urine, PMR, peritoneal fluid): up to 5 ml
- cell culture: 103–107 cells
- broth or plate bacterial or yeast culture, frozen cell pellet
- buccal, nasal, pharyngeal, vaginal, blood and saliva swabs or semen
- fresh or frozen blood: up to 1 ml
- hair: 10–30 mg
- insects: 1–30 mg
The typical efficiencies of DNA isolation from fresh biological material are given in section XIII.
50 μg DNA
- approx. 12 minutes (lysis time not included)
- 10–60 minutes for sample preparation
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 quantity of sample material and final DNA concentration expected.
Tissue: The use of 100–200 μl Elution Buffer is recommended when extracting from 2–10 mg of tissue or <104 cells. However, while extracting from 10–30 mg of tissue or 104–107 cells the volume of Elution Buffer should be increased to 200 μl.
Swab and Semen: The use of 50–100 μl Elution Buffer is recommended. The quantity of purified DNA depends on sample type and number of cells it contains; quality, however, on site’s features the swab was taken from and interindividual diversity. Usually the isolation efficiency from a single buccal swab is equal to 1–3 μg of DNA whereas, from 150 μl of semen to 2–7 μg. Bacteria and Yeast: The use of 50–100 μl Elution Buffer is recommended when extracting from no more than 109 cells (bacteria) or 108 cells (yeast). However, while extracting from a greater number of cells, the volume of elution buffer should be increased to 200 ul.
Blood: The use of 50–100 μl of Elution Buffer while extracting from 100–500 μl of blood is recommended. However, while extracting from 500–1000 μl of blood the volume of Elution Buffer should be increased to 200 ul. The quantity of purified DNA depends on the type of sample and the number of white blood cells it contains (patient’s age, his health condition, sample transport conditions, as well as storage time and method). Usually the isolation efficiency from 200 μl of blood sample from a healthy person is equal to 3–10 μg of DNA. A greater amount of DNA may be acquired from clinical samples containing increased number of white blood cells (3x 106 – 1x 107 cells/ml).
EXTRACTME GENOMIC DNA KIT (EM13) - Manual
EXTRACTME GENOMIC DNA KIT – TROUBLESHOOTING
The quality of each production batch (LOT) of the EXTRACTME GENOMIC DNA KIT is tested with the use of standard QC procedures. Purified DNA concentration and quality are evaluated by gel electrophoresis and spectrophotometer.
EXTRACTME GENOMIC DNA KIT (EM13) - MSDS
EXTRACTME GENOMIC DNA KIT (EM13) - CoA 1.2018 L. 625773
EXTRACTME GENOMIC DNA KIT (EM13) - CoA 2.2017 L.676572
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
The role of TLR9 on Leishmania amazonensis infection and its influence on intranasal LaAg vaccine efficacy
Authors Juliana Elena Silveira Pratti, Alessandra Marcia da Fonseca Martins, Juliana Paiva da Silva, Tadeu Diniz Ramos, Joyce Carvalho Pereira, Luan Firmino-Cruz, Diogo Oliveira-Maciel, Thiago Soares de Souza Vieira, Leandra Linhares Lacerda, Andre Macedo Vale, Celio G. Freire-de-Lima, Daniel C. Oliveira Gomes, Elvira M. Saraiva, Bartira Rossi-Bergmann, Herbert Leonel de Matos Guedes Products EXTRACTME GENOMIC DNA KIT (EM13) Year 2019 Source PLOS Neglected Tropical Diseases, Vol 13(2)
Analysis of bacteria associated with honeys of different geographical and botanical origin using two different identification approaches: MALDI-TOF MS and 16S rDNA PCR technique
Authors Pomastowski P, Złoch M, Rodzik A, Ligor M, Kostrzewa M, Buszewski B Products EXTRACTME GENOMIC DNA KIT (EM13) Year 2019 Source PLoS ONE 14(5)
A novel, nested, multiplex, real-time PCR for detection of bacteria and fungi in blood
Authors Tomasz Gosiewski, Danuta Jurkiewicz-Badacz, Agnieszka Sroka, Monika Brzychczy-Włoch and Małgorzata Bulanda Products EXTRACTME GENOMIC DNA KIT (EM13) Year 2014 Source BMC Microbiology 2014, 14:144
Biofilm Formation and Antimicrobial Susceptibility of Staphylococcus epidermidis Strains from a Hospital Environment
Authors Robert D. Wojtyczka, Kamila Orlewska, Małgorzata Kępa, Danuta Idzik, Arkadiusz Dziedzic, Tomasz Mularz, Michał Krawczyk, Maria Miklasińska and Tomasz J. Wąsik Products EXTRACTME GENOMIC DNA KIT (EM13), 2x PCR TaqNova-RED PCR Master Mix (RP85T), 1 Kb HyperLadder IV Year 2014 Source International Journal of Environmental Research and Public Health
In Vitro Antimicrobial Activity of Ethanolic Extract of Polish Propolis against Biofilm Forming Staphylococcus epidermidis Strains
Authors Robert D. Wojtyczka, Małgorzata Kępa, Danuta Idzik, Robert Kubina, Agata Kabała-Dzik, Arkadiusz Dziedzic, and Tomasz J.Wąsik Products 2x PCR TaqNova-RED PCR Master Mix (RP85T), EXTRACTME GENOMIC DNA KIT (EM13), 1 Kb HyperLadder IV Year 2013 Source Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2013, Article ID 590703, 11 pages
The application of genetics methods to differentiation of three Lactobacillus species of human origin
Authors Tomasz Gosiewski, Agnieszka Chmielarczyk, Magdalena Strus, Monika Brzychczy-Włoch, Piotr B. Heczko Products EXTRACTME GENOMIC DNA KIT (EM13) Year 2012 Source Ann Microbiol (2012) 62:1437–1445