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Materials for testing of RNA coronavirus, causing
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PCR Genotyping Kit
Description
PCR Genotyping Kit is designed for rapid extraction of DNA from various tissues and cells and for PCR genotyping of the extracted DNA. Kit includes DEP-25 DNA extraction Kit and Combi PPP Master Mix for Hot-Start PCR.
DEP-25 (DNA Extraction for PCR under 25 min) is a two-component reagent set for extraction of genomic DNA of various origin for PCR analysis. Extraction with DEP-25 is performed rapidly and is carried out in a single tube, mitigating chances for sample cross-contamination. The kit is amenable to high throughput-based screening and is suitable to several downstream applications such as identification of genotypes, DNA fingerprinting, and cell identity/contamination analysis. DNA can be prepared in 25 min or less and does not require lengthy enzymatic digestion, expensive column purification or phenol/chloroform extraction (Fig. 1).
Combi PPP Master Mix is dedicated for simplified hot-start PCR. It contains Taq DNA polymerase, deoxyribonucleotides, reaction buffer components, additives and monoclonal antibody anti-Taq for hot-start PCR. Samples for PCR are prepared by simple mixing 2x concentrated Combi PPP Master Mix with target-specific oligonucleotide primers, template DNA and water. Additives and the dye present in Combi PPP Master Mix allow direct loading of the PCR-amplified samples into the well in the gel without adding loading buffer. Combi PPP master mix is compatible with template DNA extracted with DEP-25 (Fig. 2).
Detailed description of the kit components, technical data, storage conditions and protocols are described in leaflets to DEP-25 DNA extraction Kit and Combi PPP Master Mix . Individual components of the kit can also be ordered separately.
Fig. 1. Schematic presentation of two methods used for preparation of genomic DNA in quality suitable for PCR analysis. Extraction with DEP-25 consists of 3 simple steps and lasts ~25 minutes. This procedure is shorter, simpler and cheaper when compared to DNA isolation using standard DNA isolation method based on proteinase K digestion and phenol/chloroform extraction.
Fig. 2. PCR amplification of genomic DNA extracted with DEP-25 (1) or isolated by standard DNA isolation method (2). For PCR with Combi PPP Master Mix, oligonucleotide primers specific for 864 bps fragment were used [Nucl. Acids Res., 36 (15):e93, 2008]. PCR amplicons were size-fractionated by agarose gel and stained with ethidium bromide. Only fragments of the expected size were visible.
Technical Data
Components and Packaging
- DEP-25 DNA Extraction Kit is supplied in bottles containing either 2x 8 ml (Cat. No. D225), 2x 30 ml (Cat. No. D226) or 2x 120 ml (D227) of START-Blue and STOP reagents.
- Combi PPP Master Mix is supplied in tubes: 500 µl (Cat. No.C208), 5x 500µl (Cat. No. C209), 25x 500µl (Cat. No. C210) + PCR Ultra H2O.
Storage
- DEP-25 DNA Extraction Kit at temperature 2 - 8°C. For short time (days) at temperature up to 35°C. This allows transport without cooling (nature friendly).
- Combi PPP Master Mix at temperature -20°C ± 5°C. For short time (days) at temperature up to 35°C. This allows transport without cooling (nature friendly).
Purity and Quality Control
- Each batch of PCR Genotyping Kit is tested for its ability to extract genomic DNA, which can be amplified by PCR. The results are verified by electrophoresis in agarose gel in the presence of ethidium bromide; only DNA band of the expected size is present (Fig. 2).
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