Because nucleic acid extraction (NAE) is the first step in many molecular biology applications, one of the first issues you must address when setting up a lab is what DNA extraction equipment you will require.
We recently evaluated various possibilities for establishing an in-house lab and gathered our findings to assist you in setting up your DNA extraction system.
Nucleic Acid Techniques & DNA extraction equipment
Different processes require various DNA extraction system available in the market. Following are some of the conventional Nucleic acid extraction techniques listed.
1. Rapid one-step extraction
This technique just has one step to use the DNA extraction system, as the name suggests. The solution is mixed with an extraction buffer comprising Tris-HCl, EDTA, sodium lauryl sarkosyl, and water-insoluble PVPP. After incubation, cooling, and dilution in double-distilled water, the combination is ready to use.
This approach requires complete DNA extraction equipment with accurate steps to get correct or satisfactory findings.
Although this technique may extract nucleic acids from non-complex materials like Gram-positive bacteria using a DNA extraction system, it is no longer utilized because PCR inhibitors are co-extracted, and buffer chemicals (such as EDTA) are carried over and can have a significant impact on downstream applications.
Furthermore, today's applications, such as qPCR and next-generation sequencing (NGS), are extremely sensitive and need high-purity nucleic acids.
2. Extraction using Chelex 100
The users need to use a proper DNA extraction system to perform these steps. An extraction buffer comprising Chelex 100 resin, SDS, NP40, and Tween 20 is added to the sample for Chelex 100 extraction.
After 30 minutes of incubation at 100°C, the mixture is centrifuged. The supernatant is removed from DNA extraction equipment and the concentration of 10 mM Tris-HCl and mM EDTA is adjusted to a final concentration of 10 mM Tris-HCl and mM EDTA.
This technique has low purity and cannot effectively remove PCR inhibitors from complex matrices due to the absence of purification stages. Furthermore, the protocol's high warmth and alkalinity can denature DNA.
As a result, Chelex 100 extraction is no longer widely utilized since other techniques, such as spin column extraction, take the same amount of time and produce far superior results.
Furthermore, traditional nucleic acid extraction techniques are ineffective for difficult-to-lyse materials (e.g., yeast, human specimens, animal tissue, or plant material), which need both chemical and physical cell lysis.
The most prevalent NAE techniques
Most laboratories today use sample- and application-specific DNA extraction equipment for spin-column or magnetic bead extraction processes to separate nucleic acids. But, before we go into these approaches in-depth, let's take a look at some solution-based methods first.
These can be beneficial for sample types that don't yield the required results when extracted using a spin column or magnetic beads, such as samples with big particles like dirt or dust, or high volume samples like chemostat cultures. The complex and highly specific Nucleic acid extraction requires an advanced DNA extraction system.
Final VerdictIn short, every nucleic acid extraction technique requires the use of complex DNA extraction equipment with knowledge and proficiency of using it.