By Dilip Kumar Arora, Surajit Das, Mesapogu Sukumar
This Springer Protocols guide is a realistic advisor to the appliance of key molecular biology strategies in microbiological examine. the point of interest is on experimental protocols, that are awarded in an easy-to-follow approach, as step by step systems for direct use within the laboratory. Notes on the right way to effectively follow the techniques are incorporated, in addition to options concerning fabrics and providers. as well as the sensible protocols, very important history details and consultant result of experiments utilizing the defined equipment are offered. Researchers in all components employing microbial platforms, akin to in molecular biology, genetics, pathology, and agricultural examine will locate this paintings of significant value.
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Extra info for Analyzing Microbes: Manual of Molecular Biology Techniques
SYBR Green) or sequence-specific probes. The procedure follows the general principle of polymerase chain reaction; its key feature is that the amplified DNA is quantified as it accumulates in the reaction in real time after each amplification cycle. Using sequence-specific primers, the relative number of copies of a particular DNA or RNA sequence can be determined. We use the term relative since this technique tends to be used to compare relative copy numbers between tissues, organisms, or different genes relative to a specific housekeeping gene (or reference gene).
An individual may have eight microsatellite repeats in one allele and—in the case of heterozygous individuals—ten repeats in the other allele. Polymorphic microsatellite loci are ideal molecular markers, which have a wide range of applications including the determination of paternity, population genetic studies, and recombination mapping. Since microsatellite repeats tend to occur in noncoding regions of the DNA, they are selectively neutral. They are also usually inherited in a co-dominant Mendelian fashion.
Random Amplified Polymorphic DNA (RAPD) fingerprinting is a low-cost yet powerful molecular typing method for organisms from bacterial species to mammals [17, 18]. RAPDs have become well-established genetic tools for genomic mapping and linkage analysis, genotype fingerprinting and identification, and quantification of genetic relationships, similarities, and variation in a variety of organisms. RAPDs are generated from genomic DNA by PCR amplification of only a small amount of total DNA using short (usually 10-mer) randomly constructed oligonucleotides.
Analyzing Microbes: Manual of Molecular Biology Techniques by Dilip Kumar Arora, Surajit Das, Mesapogu Sukumar