Enzymatic DNA/RNA Extraction Buffer
- Extracts both DNA and RNA in two simple steps
- Lyses broad spectrum of cells, without heat or mechanical agitation
- Stabilizes and protects DNA/RNA from degredation by DNase and RNase
- Large volumes of extract may be added directly to PCR reactions for sensitive detection
Fast and Efficient Lysis
The buffer incorporates two separate lysis mechanisms to maximize lysis of a wide variety of organisms. A chemical process disrupts the lipid bilayer, while enzymes simultaneously digest membrane proteins. As lysis proceeds immediately, the buffer can be used to inactivate potentially infectious agents at the time of sample collection.
Maximize DNA/RNA Recovery
RNA is notoriously susceptible to degredation by RNase, making it difficult to work with. As the buffer's specialized enzymes are active at ambient temperatures, DNase, RNase, and antimicrobial proteins are immediately digested as cells lyse, protecting DNA and RNA from degredation and increasing extraction efficiency.
Lyse Liquid or Solid Samples
The Enzymatic DNA/RNA Extraction Buffer is available in two formats. The 1X concentration allows solid samples such as swabs, cell pellets, and membrane filters to be placed into buffer for lysis.
The 10X concentration can be added directly to liquid samples such as water, biological fluids, or transport media. This minimizes dilution of the sample, allowing sensitive detection by PCR.
Simple Protocol, Direct PCR
There's no need for any centrifugation or mechanical agitation steps for most samples. Simply expose cells to lysis buffer for 15 minutes at ambient temperature, then incubate for 5 minutes at 98 ºC to inactivate enzymes. The buffer is safe to handle; all reagents are non-toxic.
The Enzymatic DNA/RNA Extraction Buffer displays low PCR inhibition, allowing larger volumes of lysate to be added to PCR reactions for more sensitive detection. With our inhibition-tolerant PCR Master Mixes, up to 25% lysate may be used depending on sample type.
The Enzymatic DNA/RNA Extraction Buffer has been tested to work with:
|Bacteria (gram-positive, gram-negative)|
|Human cells (buccal, epithelial, blood)|
|Viruses (RNA and DNA)|
The buffer degrades DNase/RNase and stabilizes nucleic acids chemically, so extracts are extremely stable short-term, even at ambient temperatures.
Both DNA and RNA extracts may be stored for up to 72 hours at 25 ºC, allowing flexibility in sample processing and transport. For long-term storage store DNA extracts at -20 ºC, and RNA extracts at -70 ºC.
|Lysis Temperature||20 – 30 ºC|
|Inactivation Temperature||98 ºC|
|Storage Conditions||-20 ºC, avoid repeated freeze/thaw cycles|
|DNA Extract Stability||72 hours at 25 ºC, long-term at -20 ºC|
|RNA Extract Stability||72 hours at 25 ºC, long-term at -70 ºC|
The Enzymatic DNA/RNA Extraction Buffer may be used license-free for commercial or diagnostic applications, and is available in bulk quantities for OEM use.
Has this been validated for SARS-CoV-2 Testing?Yes. The 10X Enzymatic DNA/RNA Extraction Buffer has been validated for SARS-CoV-2 detection from nasopharyngeal swabs in VTM media and saliva samples, while the 1X buffer has been validated for recovery from dry swabs.
How does the buffer lyse cells?
The Enzymatic DNA/RNA Extraction Buffer uses both chemical means to disrupt cell membrane lipid bilayers, and digestive enzymes to degrade membrane proteins.
The digestive enzymes also digest DNase and RNase, protecting the DNA/RNA from degredation. As the digestive enzymes are active at ambient temperatures, DNase and RNase are degraded immediately as cells lyse, maximizing nucleic acid recovery.
How does sensitivity compare with spin column purification?
Spin column purification has the advantage of concentrating the sample, but nucleic acid is lost at each purification step. While the direct PCR approach employed with the Enzymatic DNA/RNA Extraction Buffer does not concentrate the sample, it does eliminate the repeated losses of nucleic acid. Additionally, the Enzymatic DNA/RNA Extraction Buffer allows larger lysate volumes to be added to PCR reactions, aiding sensitivity.
Which approach is more sensitive for a given sample depends on the level of PCR inhibitors present and exact volumes of lysate added to the PCR reaction. The goal of the Enzymatic DNA/RNA Extraction Buffer is to provide Cq values reasonabily similar to spin column purification, but with a simpler and less costly workflow.
For Research Use Only. Not for use in diagnostic procedures.