Read e-book online Adrenergic Receptor Protocols PDF

By Curtis A. Machida

ISBN-10: 0896036022

ISBN-13: 9780896036024

Oregon health and wellbeing Sciences Univ., Beaverton. offers a suite of reproducible tools for the research of regulators of CNS-mediated habit and neural functionality. For neurobiologists, neurochemists, neurologists, and pharmacologists. DNLM: Receptors, Adrenergic - research.

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Extra info for Adrenergic Receptor Protocols

Sample text

Lefkowitz, R. , Minneman, K. , et al. (1994) IV. International Union of Pharmacology Nomenclature of Adrenoceptors. Pharmacol. Rev. 46, 121–136. 6. Johnson, R. D. and Minneman, K. P. (1987) Differentiation of α1-adrenergic receptors linked to phosphatidylinositol turnover and cyclic AMP accumulation in rat brain. Mol. Pharmacol. 31, 239–246. 7. Minneman, K. , and Abel, P. W. (1988) Comparison of α1-adrenergic receptor subtypes distinguished by chloroethylclonidine and WB4101. Mol. Pharmacol. 33, 509–514.

The sense and antisense primers should have approximately the same G + C content (40–60%). The melting temperatures, Tms, for each primer should be within 1–2°C of each other (see Note 12). 5. The primer annealing temperature for PCR is approx 5°C lower than the Tm of the oligonucleotides (see Note 13). 6. The selection of primers from known sequences can be determined visually or with computer programs. 0 (National Biosciences, Plymouth, MN) or Prime from GCG (see Note 14). 7. Primers for amplifying DNA of more than 2 kb in length (long-distance PCR, LDPCR) are designed to have higher annealing temperatures to provide greater specificity.

The reverse transcriptase enzymes most often used in these reactions are derived from Avian Myeloblastosis Virus (AMV) or Moloney Murine Leukemia Virus (M-MLV). AMV can reverse-transcribe at higher temperatures (48–55°C), thus eliminating secondary structure problems, and M-MLV can reverse-transcribe more efficiently and with less RNase H activity, but at lower temperatures (37–42°C). Other protocols are also available that utilize the same enzyme (Thermus thermophilus, Tth) for reverse transcription (70°C) as for PCR by changing the reaction conditions.

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Adrenergic Receptor Protocols by Curtis A. Machida

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