The development of non-chaotropic chemistry for nucleic acid sample preparation represents a major milestone in the history of DNA/RNA purification methods because this chemical process does not disrupt or denature the structure of macromolecules. Exclusively through non-chaotropic chemistry, the gentlest buffers may be employed, allowing for highly efficient enzymatic lysis of complex samples and for nucleic acid purification. Furthermore, the use of low salt concentrations enables the development of very fast DNA/RNA purification protocols.
Leveraging this unique and proprietary core technology, Invitek Molecular has developed several innovative sample preparation and purification solutions.
We offer a wide range of ready-to-use kits, suitable for the manual preparation of highly purified nucleic acids, as well as high-throughput purification of DNA/RNA from various starting materials on well-established laboratory devices.
Invitek Technologies
Non-Chaotropic Chemistry
Non-chaotropic chemistry is a chemical process that does not disrupt or denature the structure of macromolecules, such as proteins and nucleic acids. Ions are ranked in the so-called Hofmeister series according to their ability to denature or stabilize proteins. Protein denaturing ions (chaotropic, or “aggressive”) such as guanidinium, isothiocyante or chlorate are on one side of the spectrum while protein stabilizing ions (non-chaotropic, or “gentle”) such as Ammonium, phosphate or sulfate are on the other side.
The key advantages of non-chaotropic chemistry enable the creation of several unique solutions for sample preparation Systems.
Key Advantages
Time savings through faster protocols
Non-chaotropic conditions enable binding of nucleic acids in very low salt concentrations down to the low millimolar (mM) range. This is in sharp contrast to binding conditions required by chaotropic chemistry where 100-fold or higher concentrations in the molar (M) range are needed to achieve efficient nucleic acid binding.
Low salt concentrations reduce the number of washing steps and lead to faster protocols.
Higher DNA yields from precious samples
The non-chaotropic buffers improve reaction conditions for enzymes during sample lysis. This gentle chemical environment leads to highly efficient lysis of complex tissues as well as protein degradation by lytic enzymes. In comparison to guanidium ions (which have a strong protein denaturing effect) the non-chaotropic salts used in several of our kits actually create a stabilizing environment for lytic enzymes. The long-lasting activity of enzymes in non-chaotropic buffers leads to improved and maximized yields of DNA from complex and difficult samples.
More intact DNA
The non-chaotropic buffers provide a chemically “gentler” environment for the nucleic acids as compared to buffers based on common chaotropic salts such as guanidinium. As a consequence, DNA purified with our non-chaotropic reagent systems is often more intact with less chemical degradation – crucial attributes for optimum assay performance.
Sample Preparation System
Ready to prep
Maximizing convenience in the processing of pathogen-containing samples
The RTP range of products is based on Invitek Molecular’s unique, patent-protected non-chaotropic chemistry. RTP products greatly simplify the process of DNA/RNA purification from pathogen-containing samples.
All reagents required for DNA/RNA preparation – including lysis buffer, proteinase and/or lysozyme and carrier nucleic acid (for enhanced recovery of low amounts of target sequences) – are provided as a lyophilized master mix in a single extraction tube. This reagent set is always ready-to-use and stable at room temperature. The extraction process is initiated simply by adding a sample.
Benefits
• Lyophilized components (lysis buffer, proteinase and/or lysozyme and carrier nucleic acid) in a single extraction tube, ready-to-use, with no pre-mixing of reagents required
• Faster, more convenient DNA/RNA sample preparation via reduced handling and preparation time
• Maximum user safety
• Stable at room temperature: No refrigerator space is consumed, no need to thaw reagents
• Less waste with fewer plastic tips and vessels used
Minimal salt binding
Just two simple steps from crude sample to pure DNA
Based on Invitek Molecular’s unique and proprietary non-chaotropic chemistry the MSB range of products is the fastest way to purify DNA fragments.
The technology requires only two simple process steps:
1. DNA binding to a membrane (achieved in extremely low salt concentrations, no washing is required)
2. Elution of DNA fragments directly after binding
MSB products have been developed to purify DNA from reaction mixtures like PCR, cDNA synthesis, restriction enzyme digests or other enzymatic reactions. DNA fragments from 80 bp to 30 kbp are recovered with superior yields in a final elution volume as low as 10 µl.
Benefits
• The fastest way to purify DNA fragments from 80 bp to > 30 kbp
• Excellent purity without washing
• 80-95 % recovery rate
• Elution volume as low as 10 µl
• Less waste with fewer plastic tips and vessels used
Pre-analytical Sample Processing
Optimized sample processing for molecular diagnostics
Drawing on Invitek Molecular’s unique and proprietary non-chaotropic chemistry, the PSP product portfolio comprises an integrated sample management system: from sample collection, DNA stabilization, transportation and storage through to purification – all conveniently combined in a single kit.
The unique PSP buffer composition allows for a reliable stabilization of DNA in stool, saliva or swab samples. The stabilized samples preserve bacterial titers and can even be stored or transported at room temperature without degradation.
The purification protocol has been carefully optimized for the fast and efficient isolation of highly purified DNA from host and pathogens.
Benefits
• Reliable stabilization of host and pathogen DNA in stool, saliva or swab samples at room temperature
• Safer sample handling of bacteria with optimized pre-lysis of bacteria
• Preservation of bacterial titers at the time of sampling
• Reliable sample collection in easy-to-handle, resealable tubes
• Suitable sets of extraction kits for a variety of purposes available at Invitek Molecular
Invimag Technology
Magnetic beads for DNA/RNA sample preparation
The InviMag magnetic beads have been developed and selected for optimal performance in a variety of applications. Based on the application, sample characteristics and instrument platform used, Invitek Molecular selects the InviMag beads best suited for each particular purpose.
Characteristics
• Fast magnetic velocity via high magnetic content (70–90 % of iron oxides)
• A variety of binding capacities through different-sized beads (200 nm–1 µm)
• Excellent reproducibility and stability through uniform size distribution
• Superior suspension stability
Invitrap Technology
Selective DNA removal during RNA isolation
The unique feature of InviTrap products is the specific removal of DNA from RNA sample preparations. After sample lysis, the process has been optimized for specific binding of DNA to proprietary particles followed by removal of particle-bound DNA. The RNA is purified subsequently, leading to highly purified RNA for optimized downstream assays such as RT-PCR.
Benefits
• Highly purified RNA for better RT-PCR results
• Selective DNA removal during lysis
• No DNase digestion required
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