Introduction
The SolPure Blood DNA Kit provides a salt precipitation workflow for purification of high-molecular-weight genomic DNA from whole blood and related biological fluids. The protocol uses detergent-mediated cell lysis followed by protein precipitation and alcohol-based DNA recovery, producing genomic DNA fragments that can exceed 200 kb in size.
The scalable purification workflow allows preparation of genomic DNA suitable for PCR analysis, sequencing preparation, or DNA archiving from routine blood samples.
For silica membrane column purification laboratories may use the HiPure Blood DNA Mini Kit (D3111), while automated extraction workflows are supported by the MagPure Blood DNA Kit (D6311) and MagPure Universal DNA Kit (IVD3102).
Details
Specifications
|
Features
|
Specifications
|
|
Main Functions
|
Isolation total DNA from whole blood using economic salt out method
|
|
Applications
|
PCR, enzyme digestion, Southern hybridization,etc.
|
|
Purification technology
|
Salting out method
|
|
Process method
|
Manual (centrifugation or vacuum)
|
|
Sample type
|
Animal tissue
|
|
Sample amount
|
Unlimited
|
|
Elution volume
|
≥100μl
|
|
Time per run
|
Variation with sample amount
|
Principles
Cells are lysed with ananionic detergent in the presence of a DNA stabilizer. The DNA stabilizer limits the activity of intracellular DNases and also DNases found elsewhere in the environment. RNA is then removed by treatment with an RNA digesting enzyme. Other contaminants, such as proteins, are removed by salt precipitation. Finally, the genomic DNA is recovered by precipitation with alcohol and dissolved in Buffer TE. Purified DNA typically has an A260/A280 ratio between 1.7 and 1.9, and is up to 200 kb in size. The DNA can be safely stored at 2-8°C, -20°C, or -80°C.
Advantages
-
Safe - without phenol chloroform extraction
-
Environment friendly - the reagents used are safe, non-toxic and without pollution
-
High molecular weight - the molecular weight of genomic DNA is about 50-150kb
-
High purity - the purified DNA has A260/280=1.8-1.9, A260/230=2.0-2.5
-
Unlimited sample size - solution type operation, sample volume can be adjusted at will
-
Cost performance - the most economical nucleic acid extraction technology
Technical Validation
The SolPure Blood DNA Kit was validated using multiple blood sample types, including 1 mL human anticoagulated blood, 10 mL human anticoagulated blood, long-term stored anticoagulated blood, 5 mL porcine anticoagulated blood and pre-homogenized coagulated porcine blood. The extracted DNA showed A260/280 values of approximately 1.8–2.0, supporting reliable protein removal and DNA purity in large-volume blood DNA extraction workflows.
DNA yield increased with blood input volume and sample type. In the tested conditions, 1 mL human blood produced approximately 39 µg DNA, 10 mL human blood produced more than 300 µg DNA, and 5 mL porcine blood produced approximately 398–412 µg DNA. Stored and coagulated blood samples also produced recoverable DNA when appropriate pretreatment was used.
Agarose gel electrophoresis showed high-molecular-weight genomic DNA bands above 23 kb without obvious smearing, indicating good DNA integrity. PCR validation using human PP1 primers and porcine-specific primers further confirmed that the purified DNA was suitable for downstream amplification workflows.
Kit Contents
|
Contents
|
D331101
|
D331102
|
D331103
|
|
Purification Volumes
|
50 ml
|
300 ml
|
1000 ml
|
|
10 x Buffer RBC
|
20 ml
|
100 ml
|
3 x 100 ml
|
|
Buffer WTL
|
60 ml
|
350 ml
|
1000 ml
|
|
Buffer PPS
|
20 ml
|
120 ml
|
350 ml
|
RNase Solution
|
300 µl
|
1.8 ml
|
6 ml
|
|
Buffer TE
|
10 ml
|
30 ml
|
120 ml
|
Storage and Stability
RNase Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
