Peptide nucleic acid methods

Kristensen E. In vitro and in vivo studies on pharmacokinetics and metabolism of PNA constructs in rodents. In Peptide Nucleic Acids Methods and Protocols, Nielsen P. E. (Ed.). 2002, Humana Press (To-towa, N.J., United States) Copenhagen, pp. 259-269. 

Braasch, D. A. and Corey, D. R. (2001) Synthesis, analysis, purification, and intracellular delivery of peptide nucleic acids. Methods 23(2), 97-107. 

Nielsen P E (2005). Gene targeting using peptide nucleic acid. Methods Mol. Biol. 288 343-358. 

Peptide Nucleic Acids Methods and Protocols, edited by 178. 

PE Nielsen. In PE Nielsen, Ed. Peptide Nucleic Acids Methods and Protocols (Methods in Molecular Biology). Towana, NJ Humana Press, 2002, pp 3-26. 

An array of oligonucleotide which is composed of 20 80-mer oligos, or peptide nucleic acid probes, is synthesized either in situ (i.e., on-chip) or using conventional synthesis followed by on-chip immobilization. The resultant DNA array is then exposed to the labeled sample of DNA, hybridized, and the identity/abundance of complementary sequences determined. This method was developed at Affymetrix, Inc. and called DNA chips. Today, oligonucleotide-based chips are manufactured by many companies using alternative in situ synthesis or depositioning technologies. 

Generally, the molecular species ion is clearly detected in the case of non-volatile compounds. The method can be useful, for example, for tetrasaccharides, small peptides, nucleic acids and other organic salts, which can be detected in either the positive or negative ion mode. 

Beck F, Nielsen PE. Peptide Nucleic Acid (PNA) A DNA mimic with a pseudopeptide backbone. In Artificial DNA Methods and Applicahons. Khudyakov YE and Fields HA, eds. 2003. CRC Press, Boca Raton, FL. pp. 91-114. 

Demidov W, Kuhn H, Lavrentieva-SmoUna IV, Frank-Kamenetskii MD. Peptide nucleic acid-assisted topological labeling of duplex DNA. Methods 2001 23 123-131. 

Stender et al. (2001) have described a method based on fluorescence in situ hybridization assay (FISH) coupled with array scanner for the detection of E. coli. A Cy3-labeled peptide nucleic acid (PNA) probe complementary to a specific 16s rRNA sequence of E. coli was employed. PNA probes are DNA mimics with... 

The most common type of capture molecules used on a protein chip are antibodies, though other proteins, such as peptides, nucleic acids, enzymes and receptors, have been spotted on to chips. Protein chips can be difficult to prepare, as the proteins need to be kept in an active state, in a high concentration and in a moist environment. Typical samples investigated are cell lysates for general research or blood and urine for diagnostic applications. Fluorescence detection methods are preferred since they are simple and sensitive. Other detection methods, where labelling is not required, can take the form of SELDl or AFM. 

Various other methods, such as DNA electrotransfer, electroporation-based gene transfer, calcium phosphate nanoparticles, peptide nucleic acids, and cell penetrating peptides, round off the currently used methods in the field of nonviral gene delivery techniques. 

Recent studies using P2 have shown that it can be used to optically amplify fluorescent DNA assays [74]. The method comprises two components (a) the light harvesting luminescent conjugated polymer P2 and (b) a probe oligonucleotide consisting of a peptide nucleic acid (PNA) labeled with a reporter dye... 

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