Background

PCR is an enzyme-based in vitro method used to perform the following: 1) to amplify specific DNA fragments for molecular diagnosis of heritable disorders; 2) to screen for susceptibility to disease; 3) to identify bacterial and viral pathogens or specific target genes; 4) to biomonitor environmental changes and gene expression; 5) to study the evolutionary process at the nucleotide level. Furthermore, this technique can be used to analyze recombinant vectors to determine both the presence and orientation of the inserted DNA, determine DNA sequence, and examine gene replacement.1-1-41

Purified DNA is most often used as a template in the PCR reaction. However, it is possible to amplify specific DNA sequences without DNA purification by starting with a single living bacterial colony. This technique is known as colony PCR, or whole-cell PCR, and provides a powerful and reliable method for the rapid amplification and isolation of any gene in both gram-negative and grampositive bacteria or any gene carried on a plasmid. As an example, repetitive element sequence-based PCR (rep-PCR) with different gram-negative bacteria (e.g., Citrobacter diversus, Escherichia coli, and Pseudomonas aeruginosa) and gram-positive bacteria (e.g., Staphylococcus aureus and Streptococcus pneumoniae) were demonstrated.1-5-1

In general, broth cultures as well as colonies from agar plates and glycerol stocks can be directly utilized in the PCR mixture. To release the DNA for PCR amplification, the bacterial cell wall can be disrupted by heating. The dH2O-diluted whole-cell sample or overnight culture can also be directly applied to the PCR reaction tube with longer PCR initial denaturing time at 95 °C.

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Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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