Real-time monitoring of DNA colonies growing in a polyacrylamide gel.
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Biomedical subjects
Timur R Samatov
Publications and source records attributed to Timur R Samatov.
Expressible molecular colonies.
Carrying out polymerase chain reaction in a gel layer generates a 2-D pattern of DNA colonies comprising pure genetic clones. Here we demonstrate that transcription, translation and protein folding can be performed in the same gel. The resulting nucleoprotein colonies mimic living cells by serving as compartments in which the synthesized RNAs and proteins co-localize with their templates. Yet, due to the absence of penetration barriers, such a molecular colony display allows cloned genes to be directly tested for the encoded functions. Now, the results imply that virtually any manipulations with genes and their expression products can be accomplished in vitro.
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Molecular colony diagnostics: detection and quantitation of viral nucleic acids by in-gel PCR.
When PCR is carried out in a polyacrylamide gel, each target molecule forms a molecular colony that comprises many copies of the original template. By counting the number of colonies, one can directly determine the target titer, with 100% of the DNA molecules and approximately 15% of the RNA molecules being detected. Furthermore, because of the spatial separation of the products in the gel, no interference is observedfrom another simultaneously amplified target even if it is present at a 106 higher amount orfrom human nucleic acids that outweigh the target by up to a factor of 1,012, which is often true of clinical samples. All these features provide for an accurate and reliable assay of viruses even at very low amounts, that is, in cases most important to diagnostics.
Acrylic Resins↗