Get Peptide Nucleic Acids: Methods and Protocols PDF


By Peter E. Nielsen (auth.), Peter E. Nielsen (eds.)

ISBN-10: 0896039765

ISBN-13: 9780896039766

Peptide nucleic acid (PNA) know-how keeps to develop in significance all through molecular biology, genetic diagnostics, and molecular drugs. In Peptide Nucleic Acids: tools and Protocols, Peter Eigil Nielsen has assembled a severely evaluated number of PNA protocols which are both already good validated world wide, comparable to PCR-clamping and in situ hybridization, or that demonstrate promise of important destiny impression. uncomplicated tools for PNA oligomer synthesis and research have additionally been incorporated. Written by means of specialists with hands-on event within the tools they describe, those without problems reproducible protocols include distinctive step by step directions, tips about heading off pitfalls and on extending the tactic to different events, and introductory fabric explaining the speculation at the back of the method.
up to date and hugely sensible, Peptide Nucleic Acids: tools and Protocols presents for either amateur and skilled researchers a radical and simple advent to the optimum use of modern-day PNA know-how throughout a large choice of fields, together with molecular biology, medicinal chemistry, genetic diagnostics, and drug development.

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Additional info for Peptide Nucleic Acids: Methods and Protocols

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17. N-(2-Boc-aminoethyl)-N-(thymin-1-yl-acetyl)glycine (Boc-PNA-T monomer) (+ 97%). 18. Solutions: see Table 1. 19. Reagent A (Kaiser test): Mix 40 g phenol with10 mL water. Heat gently until all solid is dissolved. 26 M) Stability 2 mo 2 wk 1 wk 2 wk 2 wk 2 wk 2 wk 2 wk 2 wk Special requirements None Light-sensitive Light-sensitive None None None None None None water. Dilute 2 mL KCN solution with pyridine to a total volume of 100 mL. Mix the phenol and KCN/pyridine solutions. 20. 5 g ninhydrine in 50 mL ethanol (99%).

21. , Nielsen, P. , and Dahl, O. (1997) Fluoresceinconjugated lysine monomers for solid phase synthesis of fluorescent peptides. Bioconjug. Chem. 8, 503–509. Parallel Synthesis of Conjugate Libraries 53 4 Parallel Synthesis of PNA-Peptide Conjugate Libraries Satish Kumar Awasthi and Peter E. Nielsen 1. Introduction For a large variety of studies, it would be advantageous to be able to synthesize a library of individual peptide nucleic acid (PNA) oligomers in parallel. This is especially useful for screening target positions for antisense reagents along the mRNA (gene walk) as optimal target sites have to be determined emperically to a very large degree.

3. 4. Swell 50 mg of downloaded PNA resin in DCM overnight. Deprotect resin using TFA/DCM/anisole (50/45/5) (2 × 3 min). Wash with DCM, DMF, DIEA in DCM (5/95), DMF. 7 mg dissolved in 200 µL NMP), HBTU (36 mg dissolved in 200 µL NMP) and NMM (22 µL). All three solutions were mixed and allowed to preactivate for one min, after which the mixture was added to the resin and allowed to couple for 30 min under gentle mixing. 5. Wash the resin using DMF, DCM. 6. 2. If the test is positive, repeat coupling (see step 4).

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Peptide Nucleic Acids: Methods and Protocols by Peter E. Nielsen (auth.), Peter E. Nielsen (eds.)

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