How Size Selection Works

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How Size Selection Works 
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How Just-a-Plate or Just-a-Tube Size-selected PCR Fragment Purification Kit Works


Just-a-Plate or Just-a-Tube  Size-Selection PCR Fragment Purification Kit is designed and optimized for simple and purification of size-selected PCR products directly from PCR or after PCR normalizaton in NGS sample prep workflow. DNA samples can be simultaneously purified in a regular 2 ML tube or a 96-well microplate format in less than 30 minutes. After PCR reaction or PCR normalization, PCR products can be purified with high efficiency. No spin-column, filter plate, silica membrane or magnetic beads are needed for the isolation and no vacuum and filtration are required in the process.  Based on Charm Biotech developed Solid Surface Reversible Binding (SSRB) technology, Just-a-Tube or Just-a-Plate system employs the regular 2 ml tube or 96-well plate surface coated with proprietary turbo-binders as selective nucleic acid capturers to efficiently bind PCR DNA from reaction mixtures. If 400 bps - 700 bps DNA fragments need to be isolated,  at first, DNA fragments longer than 700 bps, such as DNA template are attached to the wall of  the 1st binding tube (or plate) after the 1st binding buffer added. DNA fragments shorter than 700 bps will remain in the solution. After the solution is transferred to the 2nd binding tube (plate), 2nd binding buffer is added to the solution,  the target 400 bps-700 bps DNA,  can specifically interact with the turbo binder of the 2nd binding tube (or plate) in the extraction buffer, while primers, nucleotides, adapters and other short failure PCR products are not bound and remain in the solution. After washing the 2nd binding tube (plate) with washing buffer to remove unbound material, the purified 400bps-700 bps PCR prodcuts can be easily eluted in 10 mM Tris Elution Buffer. Without using vacuum, or magnetic device,  the unique centrifugation-based procedure of the kit reduces opportunities for cross-contamination between wells and makes the precedure easy to perform. The purified and normalized size-selected PCR fragments are ideal for use in NGS, such as Illumina MiSeq sequencing.


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