New PDF release: Analyzing Microbes: Manual of Molecular Biology Techniques

By Dilip Kumar Arora, Surajit Das, Mesapogu Sukumar

ISBN-10: 3642344097

ISBN-13: 9783642344091

This Springer Protocols guide is a pragmatic consultant to the appliance of key molecular biology concepts in microbiological examine. the point of interest is on experimental protocols, that are offered in an easy-to-follow method, as step by step tactics for direct use within the laboratory. Notes on how one can effectively practice the strategies are incorporated, in addition to suggestions relating to fabrics and providers. as well as the sensible protocols, vital heritage details and consultant result of experiments utilizing the defined equipment are provided. Researchers in all parts using microbial structures, comparable to in molecular biology, genetics, pathology, and agricultural examine will locate this paintings of serious value.

Show description

Read Online or Download Analyzing Microbes: Manual of Molecular Biology Techniques PDF

Similar microbiology books

Troubleshooting the Sequencing Batch Reactor (Wastewater by Michael H. Gerardi PDF

The sensible advisor on what to do correct whilst organic impacts reason a sequencing batch reactor to head mistaken This richly illustrated, effortless advisor contains forth the legacy demonstrated via earlier variants within the Wiley Wastewater Microbiology sequence by way of focusing realization at the combined amassing of organisms cohabitating inside of a sequencing batching reactor (SBR), and the foremost roles their biology performs during this wastewater processing tank's functionality.

The Trichomycetes: Fungal Associates of Arthropods by Robert W. Lichtwardt PDF

Institutions and interactions among species of organisms are phenomena shared via all residing issues. What varies is the level to which the extra long-lasting interactions are invaluable or damaging to a given species and the measure of intimacy and reliance which one organism could have constructed in organization with one other.

Download e-book for kindle: Outbreak: Cases in the Real-World Microbiology by Rodney P. Anderson

"Outbreak: situations in Real-World Microbiology is an interesting new textbook aimed toward introductory microbiology teachers and scholars. Generously illustrated with colour pictures, the booklet integrates headline-making disorder outbreaks into the microbiology coursework and is the 1st paintings of its type to make use of fabric that at once affects college-age scholars.

Additional info for Analyzing Microbes: Manual of Molecular Biology Techniques

Sample text

SYBER Green binds double stranded DNA and upon excitation emits light. An increase in DNA product during PCR leads to an increase in fluorescence intensity and is measured at each cycle, thus allowing DNA concentrations to be quantified (Fig. 4). The advantages of SYBER Green are that it is inexpensive, easy to use, and sensitive. However, dsDNA dyes such as SYBR Green binds to all dsDNA PCR products, including nonspecific PCR products (such as “primer dimers”). This can potentially interfere with/or prevent accurate quantification of the intended target sequence.

Denaturation at 95  C for 15 s. 2. Annealing/extension at 60  C for 1 min (repeated 40 times). 4. 1. 2. Probe Selection Criteria Whenever possible, primers and probes should be selected in a region with a G/C content of 30–80 %. Regions with G/C content in excess may not denature well during thermal cycling, leading to a less efficient reaction. In addition, G/C-rich sequences are susceptible to nonspecific interactions that may reduce reaction efficiency and produce nonspecific signal in SYBR Green assays.

The more times the denaturing and annealing takes place, the more opportunities there are for the TaqMan probe to bind and, in turn, the more emitted light is detected, and its geometric increase corresponding to exponential increase of the product is used to determine the threshold cycle (Ct) in each reaction. Fluorescent reporter probes are more accurate and reliable of the method. It uses a sequence-specific RNA or DNA-based probe to quantify only the DNA containing the probe sequence; 26 Praveen and Koundal therefore, use of the reporter probe significantly increases specificity, and allows quantification even in the presence of some nonspecific DNA amplification.

Download PDF sample

Analyzing Microbes: Manual of Molecular Biology Techniques by Dilip Kumar Arora, Surajit Das, Mesapogu Sukumar


by Christopher
4.0

Rated 4.19 of 5 – based on 34 votes