AUTOMATABLE AND RAPID DNA/RNA EXTRACTION
FROM HUMAN WHOLE BLOOD USING SILICA-BASED MATRIX
Procedure for purifying DNA from different biological sources using a silica-based method developed by ISENET.
UNIQUE FEATURES OF THE PROTOCOL:
1. Soluzione G stabilised Nucleic acid in whole blood and buffy-coat facilitating blood transportation at Room Temperature;
2. Scalable procedure use 50-1000μL of whole blood;
3. Rapid: it can extract nucleic acids from 24 samples in less than 1h;
4. Yields (30-50μg);
5. Pure and high-quality nucleic acids.
QUALITY CONTROL PERFORMANCE:
Total DNA extracted from whole blood and electrophoresed through a 0,8% agarose gel stained with ethidum bromide.
The samples are quantified by Nanodrop ND-1000 UV-VIS Spectrophotometer (NanoDrop Technologies).
DNA AMPLIFICATION WITH STANDARD PCR:
Randomly selected DNA products with the following sample codes:THCB1697, THCB1566, THCB1327, THCB1394, THCB1213, THCB1026 and THCB1666 were tested for PCR.
RNA QUALITY CONTROL (QC) TESTING:
(A) Total RNA extracted from blood and electrophoresed through a 1,2% formaldehyde agarose gel stained with ethidium bromide. (B) Electropherogram peaks obtained by capillary electrophoresis on 2100 Bioanalyzer (Agilent Technologies) testing RNA integrity.
(C) Spectophotometric UV scan image obtained when testing RNA purity and concentration by a scanning UV measurement (Nanodrop spectrophotometer).