PCR kit ALPREP for DNA/RNA extraction from biological material by magnetic sorption method
The ALPREP kit is configured for the simultaneous DNA&RNA manual or automatic extraction from biological material. This step is followed by DNA/RNA preparation analysis with the help of polymerase chain reaction (PCR) and polymerase chain reaction with a reverse transcription step (RT-PCR).
The functional purpose of the ALPREP reagent kit is to provide the preanalytical stage for the biological samples subsequent analysis, performed during the clinical laboratory diagnostics of human infectious pathology using PCR or RT-PCR. The kit allows to obtain a DNA/RNA samples free of RT-PCR and PCR inhibitors, which provides high analytical capabilities for subsequent analysis.
Field of application. Professional use for research purposes, in specialized medical & healthcare institutions.
Specimens. Extraction procedure is performed for biological material (clinical samples), including swabs, scrapings, sputum, white blood cells, urine (sediment), ejaculate, prostatic fluid (100 to 10₆ cells per sample).
The volume of the test sample is 100 μl.
The total time of the procedure (DNA/RNA extraction from the single sample):
- 40 minutes for manual method
- 35 minutes for automatic method
ALPREP reagent kit is used at the preanalytical stage of the analysis of clinical samples.
Purity of isolated DNA/RNA (A260/A280 ratio) is at least 1.7 for biological samples.
The efficiency of DNA/RNA isolation is ranged from 30 up to 70% (at least 30%).
The obtained purified DNA/RNA sample can be used for subsequent analysis by PCR, RT-PCR and their variations.
ALPREP reagents kit is based on the reversible binding of nucleic acid molecules with the surface of magnetic sorbent particles. The sample is treated with a lysis buffer in the presence of magnetic sorbent particles (magnetic beads). As a result, cell membranes, viral membranes and other biopolymer complexes are destroyed and nucleic acids are released. Dissolved nucleic acid binds to the sorbent particles, while other components of the lysed biological material remain in the solution and are removed after magnetic sorbent precipitation on a magnetic rack followed by several washing steps. When an elution buffer is added to magnetic sorbent, nucleic acids are eluted from the surface of the sorbent into solution, which is then separated from the sorbent particles by magnetic force. As a result of this procedure, a highly purified DNA (RNA) sample is obtained, free of PCR-inhibitors, which ensures high analytical sensitivity of the PCR study.