What is the Solid Surface Reversible Binding (SSRB) Technology ?
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Solid Surface Reversible Binding (SSRB) technology is a novel , patent-pending solid surface-based technology that is designed to bind nucleic acid on the surface of solid materials, such as microplates, tubes and beads. The current system utilized a 96-well microplate or tubes coated with proprietary turbo-binders acting to selectively capture and efficiently bind nucleic acids from a variety of cell lysates including bacteria, yeast,
plant or animal tissues, blood cells, cultured cells, body fluids and FFPE samples. In the presence of binding buffers, nucleic acid specifically interacts with the turbo binders and binds to the surface of a microplate well or a tube while proteins and other contaminants remain in the solution. Unbound materials is removed in a washing step. The purified nucleic acids, such as plasmid DNA, gDNA, total
RNA, mRNA, microRNA and viral RNA and DNA , can easily be eluted in 10 mM Tris elution buffer or water. The detailed procedure is listed below.
Step 1: Nucleic Acid
Capture: Tissue cells are lysed and crude nucleic acids are captured by the
turbo-binder on the surface of the solid surface through a mediator-assisted
interaction, leaving cell debris and protein behind in solution.
Step 2: Contaminant Removal:
Any residual protein and cell debris are washed away to leave pure intact
nucleic acid captured on the solid surface. Mediators also are washed away in
the process.
Step
3: Purified Nucleic Acid Elution: In an aqueous condition, after mediators are removed during the washing step, nucleic acid can be separated from the turbo-binder
and pure nucleic acid is eluted in the water or Tris buffer, ready for use in
any downstream applications.
The greatest feature of this technology over other nucleic acid purification technologies, such as silica membrane or magnetic beads methods, is its highest recovery rate of sample nucleic acids from a biological sample (please see below). So the method is best suitable for nucleic acid isolation from samples with limited resources, such as clinic patients samples, LCM samples, NGS samples etc. When your original source sample
nucleic acid is in a very limited (low) quantities, you can try our Just-a-Plate or Just-a-Tube method, to isolate the nucleic acids with maximum recovery rate over other spin-column and magnetic beads methods.
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Why Just-a-Plate technology is better than
conventional membrane (spin-column) or magnetic bead based methods for automation?
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Traditionally, almost all technologies used in the automatic
nucleic acid purification process involve the usage of silica membrane or magnetic
beads in multiple-well plate format. In this way, either magnetic force or
vacuum device have to be employed in the process, these add the technical
difficulty both in the hardware design and in the sample processing steps,
making the purification less flexible and higher cost.
Also, It is conceivable that residual buffer, located in spaces between the beads, or between the silica membrane and its holding plastic devices,
would be difficult to remove. Buffer exchange in the washing step is often incomplete, resulting in the
contamination of ethanol or other organic reagents in the final purified
sample, which will interfere with the downstream applications. To solve the
problem, long period vacuum or “drying” processing has to be employed before
the final elution step to allow completely “drying” of the membrane or magnetic
beads. This will make the extraction
procedure extremely tedious and time-consuming.
Further, membrane
and bead extraction methods typically lose 30% -50% of the nucleic acid due to
insufficient binding, incomplete elution and dead space inherited with membrane
and beads. That is why we usually collect only 50 µl –70 µl of final
elute even we apply 100 µl elution buffer in the final sample elution step in
automation extraction process. In most case, the amount of eluted nucleic acid is enough for the
downstream application. But if your sample amount is limited and there is no
room for any sample loss, Just-a-Plate kit is your right choice to limit your
sample loss and to generate the maximum recovery.
In
contrast to the membrane and beads based plate methods, Just-a-plate kit is a
non-membrane and non-bead based extraction system. Based on charm biotech
developed novel Solid Surface Reversible Binding (SSRB) technology
(patent pending),
Just-a-plate system employs the 96-well plate surface coated with proprietary
turbo-binders as selective nucleic acid capturer to efficiently extract nucleic
acids from a variety of sample resources. Every steps, including nucleic acid
binding, solution washing, buffer exchanging and sample releasing, all occur on
the surface of the plate. There are no dead space and sample trapping on the
plate solid surface. This unique solid-surface capturing method eliminates
problems related with membrane and beads, such as less flexibility, high cost,
loss of sample, sample contamination with organic solvent, tedious and
time-consuming procedure as we explained above. The Just-a-Plate system allows
for maximum high-quality nucleic acid recovery in an automation friendly
procedure with efficient sample capturing, easy and efficient buffer washing
and completely and fast sample elution.
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