Independent control of histopathology, tumor cell content of routine pathological specimens, and the selection of tissue areas for tissue extraction were determined by the pathologist. Subsequently, cylinders 1.5 mm in length and 2 mm in height were stamped out from the formalin-fixed and paraffin-embedded (FFPE) tissue blocks using an 18 Charrière core stamp and subjected to DNA isolation. Genomic DNA was extracted using the automated MagNA Pure LC 2.0 system and MagNA Pure LC DNA isolation kit II - tissue (Roche Diagnostics Deutschland, Roche Applied Science, Mannheim, Germany). The quality of extracted DNA was assessed using spectrophotometry, gel electrophoresis, and quantitative PCR, which characterized the yield, purity, and grade of degradation of isolated DNA. Bisulfite conversion of DNA was carried out using the EZ DNA Methylation-Gold Kit (Zymo Research Corporation, Irvine CA, USA) and 1 µg of isolated DNA. Fully methylated and converted DNA, as well as unmethylated bisulfite-converted DNA controls, were used as reported previously [21] (link).
Magna pure lc dna isolation kit 2 tissue
Manufactured by Roche
Sourced in Germany
The MagNA Pure LC DNA isolation kit II - tissue is a laboratory equipment product designed for the automated extraction and purification of DNA from various tissue samples. It utilizes magnetic bead-based technology to efficiently isolate DNA, enabling consistent and reliable results for downstream applications.
Automatically generated - may contain errors
2 protocols using magna pure lc dna isolation kit 2 tissue
1
Tissue Extraction and DNA Isolation from FFPE Samples
2
DNA Isolation and Real-Time PCR for Food Products
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For DNA isolation, 50 mg of food products and control samples were taken. Then lysis and purification with chloroform were carried out using the reagents of the Sorb-GMO-B Kit (Syntol, Moscow, Russia) according to the manufacturer's instructions. Further DNA isolation was performed at the MagNA Pure LC 2.0 isolation station (Roche) using the MagNa Pure LC DNA Isolation Kit II (Tissue) (Roche, Mannheim, Germany) (Kurbakov et al., 2019). Real-time PCR was performed on an ANK-32 amplifier (Syntol, Moscow, Russia). The reaction mixture (Syntol, Moscow, Russia), with a volume of 30 μL, contained primers with a concentration of 300 nM, a probe with a concentration of 150 nM, 2.5 mM MgCl2, dNTP with a concentration of 0.25 mM each, SynTaq polymerase with a concentration of 2.5 activity units, and 5 μL of isolated DNA. The PCR reaction mode was as follows: preliminary denaturation at 95 °C, 7 min, and 35 amplification cycles (60 °C for 40 s and 95 °C for 15 s).
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