-
Quick and reliable method to analyze meiotic segregation patterns in Coprinus cinereus using the polymerase chain reaction
Time of Update: 2021-02-18
The following components were added to the indicated concentrations after the first denaturation step for PCR-amplification (final volume 100 microliters): 67 mM Tris-HCl (pH 8.8), 16 mM (NH4)2SO4, 10 mM 2-mercaptoethanol, 3 mM MgCl2, 2 glycerol, 1 micromole of each primer (see below), dATP, dCTP, dGTP (each 200 uM), dUTP (500 uM), and 2.5 units of Amplitaq DNA Polymerase (Perkin Elmer Cetus).
-
Analysis of Human Cytomegalovirus Using the Polymerase Chain Reaction
Time of Update: 2021-02-17
Similarly, reverse transcription-PCR ( RT-PCR ) allows the study of targeted gene expression, by reverse transcription of RNA to complementary DNA ( cDNA ), followed by amplification of target DNA using predetermined primers.
-
The Detection of Enteroviruses in Water and Associated Materials Using the Polymerase Chain Reaction
Time of Update: 2021-02-01
Although not a cause of gastroenteritis, enteroviruses may cause disease including paralysis, meningitis, myocarditis, and cardiomyopathy, and less severe infections, such as, colds and fever, mainly in young children, They are therefore a public health concern, and in order to make proper evaluation of their significance, in water techniques are required that are rapid and reliable and can accommodate large numbers of samples in one test batch.
-
Use of Fimers to Eliminate Polymerase Chain Reaction and Primer-Dimer Artifacts and to Increase Yield in BAC-Sequencing Reactions
Time of Update: 2021-01-27
Ideally, under the elevated temperatures used in a typical amplification, the primers should hybridize only to the target sequence.
However, there is a relatively narrow range of conditions (temperature, concentrations of ions and denaturing agents) for specific annealing of an oligonucleotide to its complementary target, and often it does not coincide with other requirements of the cycle-sequencing reaction.
-
Direct DNA Sequencing of Polymerase Chain Reaction Products Using Magnetic Beads
Time of Update: 2020-12-08
The use of magnetic particles in many fields of biochemistry, molecular biology, and medicine has been well documented and several magnetic particles are now available for diagnostic and cell separation purposes.
-
Homogenous Detection of Nucleic Acids Using Self-Quenched Polymerase Chain Reaction Primers Labeled With a Single Fluorophore (LUX Primers)
Time of Update: 2020-11-27
This design provides a low initial fluorescence of the primers that increases upon formation of the PCR product.
The hairpin oligonucleotides (ΔG from -1.6 to -5.8 kcal/mol) are as efficient as linear primers and provide additional specificity to the PCR by preventing primer-dimers and mispriming.