Basic guides to PCR, labeling neurons featured in Cold Spring Harbor Protocols|
COLD SPRING HARBOR, N.Y. (Wednesday, April 1, 2009) – Along with new cutting-edge methods, Cold Spring Harbor Protocols is home to an in-depth library of basic laboratory methods. The April issue (www.cshprotocols.org/TOCs/toc4_09.dtl) features two of these standard techniques.
From molecular biology researchers to law enforcement forensics laboratories, polymerase chain reaction (PCR) is the commonly used method for nucleic acid amplification. But PCR is often difficult to optimize, and failure to do so can lead to undefined and unwanted products, or a complete lack of amplification altogether. To help avoid these issues, Kenneth Roux from Florida State University (http://bio.fsu.edu/~roux/) provides Optimization and Troubleshooting in PCR. The article addresses various optimization strategies including touchdown PCR and hot-start PCR. Magnesium concentration, buffer pH, and cycling conditions are also considered. The article is freely accessible on the website for Cold Spring Harbor Protocols (http://cshprotocols.cshlp.org/cgi/content/full/2009/4/pdb.ip66).
The loading of individual cells with fluorescent probes via patch pipettes is regularly performed in neuroscience research laboratories. This method allows for combined electrophysiological and optical measurements at a quantitative level. Arthur Konnerth and colleagues from the Institut fÜr Physiologie der Ludwig-Maximilians-Universität München (http://www.imprs-ls.de/professors/konnerth.htm) have written Dye Loading with Patch Pipettes, available in the April issue. This patch-clamp methodology has been successful for single-cell dye labeling in cultured neurons, brain slices, and in vivo preparations. A wide range of dyes can be loaded using this method, including probes for morphological reconstruction, ion-sensitive indicator dyes for monitoring second-messenger cascades, and dye-labeled proteins for fluorescence resonance energy transfer (FRET), fluorescence correlation spectroscopy (FCS), and fluorescence recovery after photobleaching (FRAP) studies. The most widespread application of this technique has been for Ca2+ imaging. This method is freely accessible on the website for Cold Spring Harbor Protocols (http://cshprotocols.cshlp.org/cgi/content/full/2009/4/pdb.prot5201).
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About Cold Spring Harbor Protocols:
Cold Spring Harbor Protocols (www.cshprotocols.org) is a monthly peer-reviewed journal of methods used in a wide range of biology laboratories. It is structured to be highly interactive, with each protocol cross-linked to related methods, descriptive information panels, and illustrative material to maximize the total information available to investigators. Each protocol is clearly presented and designed for easy use at the bench—complete with reagents, equipment, and recipe lists. Life science researchers can access the entire collection via institutional site licenses, and can add their suggestions and comments to further refine the techniques.
About Cold Spring Harbor Laboratory Press:
Cold Spring Harbor Laboratory Press is an internationally renowned publisher of books, journals, and electronic media, located on Long Island, New York. Since 1933, it has furthered the advance and spread of scientific knowledge in all areas of genetics and molecular biology, including cancer biology, plant science, bioinformatics, and neurobiology. It is a division of Cold Spring Harbor Laboratory, an innovator in life science research and the education of scientists, students, and the public. For more information, visit www.cshlpress.com.
For content and submission information:
David Crotty (firstname.lastname@example.org; 516-422-4007), Executive Editor, Cold Spring Harbor Protocols
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Stephanie Novara (novara.cshl.edu; 516-422-4159), Journals Marketing Manager, CSHL Press