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![]() | EM13-250 | 250 extractions | Ask for an offer |
![]() | EM13-050 | 50 extractions | Ask for an offer |
PRINCIPLE
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.
PRODUCT SPECIFICATIONS
SAMPLE MATERIAL
- 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
EFFICIENCY
The typical efficiencies of DNA isolation from fresh biological material are given in section XIII.
BINDING CAPACITY
50 μg DNA
TIME REQUIRED
- approx. 12 minutes (lysis time not included)
- 10–60 minutes for sample preparation
DNA PURITY
A260/A280 ratio = 1.7 – 1.9
RECOMMENDATIONS AND IMPORTANT NOTES
DNA elution
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) – Discontinued - Manual
EXTRACTME GENOMIC DNA KIT – TROUBLESHOOTING
QUALITY CONTROL
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) – Discontinued - MSDS
EXTRACTME GENOMIC DNA KIT (EM13) – Discontinued - CoA 1.2018 L. 625773
EXTRACTME GENOMIC DNA KIT (EM13) – Discontinued - CoA 2.2017 L.676572
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White pupae phenotype of tephritids is caused by parallel mutations of a MFS transporter.
Authors Christopher M. Ward, Roswitha A. Aumann, Mark A. Whitehead, Katerina Nikolouli, Gary Leveque, Georgia Gouvi, Elisabeth Fung, Sarah J. Reiling, Haig Djambazian, Margaret A. Hughes, Sam Whiteford, Carlos Caceres-Barrios, Thu N. M. Nguyen, Amanda Choo, Peter Crisp, Sheina B. Sim, Scott M. Geib, František Marec, Irina Häcker, Jiannis Ragoussis, Alistair C. Darby, Kostas Bourtzis, Simon W. Baxter & Marc F. Schetelig Products EXTRACTME GENOMIC DNA KIT (EM13) – Discontinued Year 2021 Source Nature Communications volume 12, Article number: 491 (2021) -
Gene Sequencing and Phylogenetic Analysis: Powerful Tools for an Improved Diagnosis of Fish Mycobacteriosis Caused by Mycobacterium fortuitum Group Members
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New endosymbiotic relationships between Mucoromycota and Burkholderiaceae representatives
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Linezolid-resistant Enterococcus faecium strains isolated from one hospital in Poland –commensals or hospital-adapted pathogens?
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Electrochemical Immunosensors Based on Screen-Printed Gold and Glassy Carbon Electrodes: Comparison of Performance for Respiratory Syncytial Virus Detection
Authors Białobrzeska W., Firganek D., Czerkies M., Lipniacki T., Skwarecka M., Dziąbowska K., Cebula Z., Malinowska N., Bigus D., Bięga E., Pyrć K., Pala K., Żołędowska S., Nidzworski D. Products EXTRACTME GENOMIC DNA KIT (EM13) – Discontinued, EXTRACTME VIRAL RNA KIT (EM39) – Discontinued Year 2020 Source Biosensors. 2020; 10(11):175 -
Investigating the Role of Methylation in Silencing of VDR Gene Expression in Normal Cells during Hematopoiesis and in Their Leukemic Counterparts
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Increased photoinactivation stress tolerance of Streptococcus agalactiae upon consecutive sublethal phototreatments
Authors Michal Pieranski, Izabela Sitkiewicz, Mariusz Grinholc Products EXTRACTME GENOMIC DNA KIT (EM13) – Discontinued Year 2020 Source Free Radical Biology and Medicine, Volume 160, 2020, Pages 657-669 -
Identification of Entomopathogenic Fungi as Naturally Occurring Enemies of the Invasive Oak Lace Bug, Corythucha arcuata (Say)(Hemiptera: Tingidae)
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The New Klebsiella pneumoniae ST152 Variants with Hypermucoviscous Phenotype Isolated from Renal Transplant Recipients with Asymptomatic Bacteriuria—Genetic Characteristics by WGS
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Laccase activity of the ascomycete fungus Nectriella pironii and innovative strategies for its production on leaf litter of an urban park
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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) – Discontinued, EXTRACTME DNA GEL-OUT KIT (EM08) – Discontinued Year 2019 Source Southeast Europe Journal of Soft Computing, 8, March 2019 -
Association of ABCB4 and ABCB11 nucleotide variants with intrahepatic cholestasis of pregnancy
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The role of TLR9 on Leishmania amazonensis infection and its influence on intranasal LaAg vaccine efficacy
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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) – Discontinued Year 2019 Source PLoS ONE 14(5) -
A novel, nested, multiplex, real-time PCR for detection of bacteria and fungi in blood
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Biofilm Formation and Antimicrobial Susceptibility of Staphylococcus epidermidis Strains from a Hospital Environment
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In Vitro Antimicrobial Activity of Ethanolic Extract of Polish Propolis against Biofilm Forming Staphylococcus epidermidis Strains
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The application of genetics methods to differentiation of three Lactobacillus species of human origin
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