Injection polymerase chain reaction

Biodistribution analysis is conducted at the molecular level. The current gold standard is a quantitative polymerase chain reaction (Q-PCR) assay that detects the number of vector copies per microgram of genomic DNA. Biological fluids and tissue samples are carefully harvested (to avoid crosscontamination) from control and gene therapy product-injected animals at... 

DHPLC is usually performed on a styrene-divinylbenzene-based polymeric stationary phase, with a mobile phase that contains triethylammonium acetate as the IPR to provide adequate reversed phase (RP) retention for the negatively charged nucleic acid molecules. The samples are usually amplified according to polymerase chain reaction (PCR) protocols and then injected into the chromatographic system. 

Biodistribution studies are conducted to evaluate the dissemination and persistence of nucleic acid/DNA and viral vector-based vaccines. Vector persistence should be examined at more than one time point up to at least 21 days or more after the last injection 22 unpublished data) and the biodistribution and persistence of a plasmid is usually dependent upon the route of administration.20 These studies can be combined with a general toxicology study or may be conducted separately. In most cases, the DNA is extracted from the selected tissues and assessed by quantitative Polymerase Chain Reaction (qPCR).1516... 

Polymerase chain reaction (PCR) and reverse-transcription-PCR (RT-PCR) analysis demonstrated the presence of Ad5 DNA and FGF-4 mRNA, respectively, in the injected pig hearts [51]. Immunoblotting showed that pigs treated with... 

In some experiments, it may be necessary to recover microinjected cells (e.g., for electrophoresis, polymerase chain reaction). One way to do this is to plate cells on small pieces of coverslips placed inside a culture dish, inject all cells on pieces of coverslips and then remove the pieces with forceps. 

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