http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-2017058344-A1
Outgoing Links
| Predicate | Object |
|---|---|
| assignee | http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_63e215336df25fd371cfe6a5e39cc2ee |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6806 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6874 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N27-44791 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N33-48721 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N27-4473 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N27-447 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12Q1-68 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N33-487 |
| filingDate | 2016-08-31-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| inventor | http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_491e30294a0a1a620839a1391466dd8f |
| publicationDate | 2017-03-02-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | US-2017058344-A1 |
| titleOfInvention | Mapping protein bound regions on dsDNA by reaction of unblocked Thymidines with Osmium tetroxide 2,2'-bipyridine. |
| abstract | Materials, methods, and systems for determining the position, i.e. the sequence within a double stranded nucleic acid, where a protein is bound are disclosed and described. Materials can include dsDNA, dsRNA, and dsDNA-mimics. Materials are first reacted with Osmium tetroxide 2,2′-bipyridine to partially osmylate the nucleic acid (osmylated or labeled polymer) and monitored by HPLC or CE in order to stop the labeling before it reaches 50%. This preserves the double stranded structure of the nucleic acid and leaves the protein bound to it during the labeling. Labeling occurs randomly at the whole length of the polymer but leaves the region where the protein binds intact. Methods are provided to describe preparation of the osmylated polymers, their purification, and characterization. Labeled polymers may be subject to voltage-driven translocation via nanopores of appropriate width so that the polymer can traverse. There are two choices: Either use nanopores suitable for dsDNA, or use nanopores suitable for ssDNA and denature the labeled polymer before testing. Either way the translocation is monitored and reported as a current vs. time (i-t) profile. The current is stable, but fluctuates during the polymer's translocation in a manner that pinpoints the region where characteristically no osmylated base appears (less current reduction in i-t), while the majority of the trace will exhibit a pattern corresponding to the osmylated bases interspersed among the intact bases (more current reduction in i-t). The region where the protein is bound can be inferred by comparison of the i-t data of the two osmylated polymers (with or without bound protein). |
| priorityDate | 2015-08-31-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
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