Taq polymerase

PCR technique for both polymorphisms was performed in 2µl genomic DNA, 10 µl buffer (pH 8.4) 0.8 µl MgCl2, 0.7 µl dNTPs, 1 µl of each primer and 0.5 U Taq DNA polymerase. Then 5µl of products were incubated with 0.4U of Taq1 restriction enzyme at 65 °C for 48 h. Finally the products of the Taq1B digestion were electrophoresed on 1.5% agarose gel. For -629C/A polymorphism, 5µl of PCR products were incubated with 0.4U of Van91I restriction enzyme at 37 °C for 48 h. The thermocycler conditions after

especially agriculture and medicine. Humans have been genetically modifying plants and animals for thousands of years through selective breeding, however the direct manipulation of DNA has only existed since the 1970s. One such manipulation of DNA is Polymerase chain reaction, or PCR, a technique used generate millions of copies of a DNA segment from one pre existing DNA segment. However PCR does not alter the DNA of an organism, it replicates it. PCR finds its main uses in medicine,

Amplification of 16S Ribosomal DNA Sequences using Polymerase Chain Reaction Edwina Abou Haidar, Houssam Al Koussa, Mary AbedAlAhad. Department of Biology, Lebanese American University, Byblos, Lebanon Abstract The 16s rRNA gene sequencing is a widely common amplicon sequencing method used to identify and compare bacteria in a given sample. This method is well established and allows to study phylogeny and taxonomy of complex microbiomes. In this study, an unknown sample of extracted microbial

GCT TGC CTT TCA GGT CTG – 3’ For primers F4/R2 and B4/B5 amplification reaction mixture was prepared in a volume of 25 µl containing 1X PCR buffer, 1.5 mM MgCl2, 200 µM each deoxynucleoside triphosphate, 20 pmol each primer, 1 U of Taq DNA polymerase and 3 µl of template DNA (~100ng). The following PCR conditions were used (Table 2). Table 2: Brucella PCR Program Stage Step Temperature °C (for F4/R2) primers Duration (for F4/R2) primers Temperature °C (for B4/B5) primers Duration (for

strong base and heated to 95 degrees Celsius, which leads to breaking down the hydrogen bonds between the nitrogenous (Tracey, 2017). Once all the previous steps have been completed, the PCR reaction can commence after mixing with primers, Taq DNA polymerase, reaction buffers and dNTPS. The Thermocycler was utilized to facilitate PCR making it faster, more reliable and cheaper (Tracey, 2017). The sample will undergo 35 amplification cycles, producing 3.44x1010 target sequences [ 235= 3.44x1010

Pneumonia is a very common infectious disease that causes substantial morbidity and mortality.“It is the leading cause of death in children worldwide”.8 Prevention measures such as immunization, proper nutrition and addressing environmental factors in addition with disease management according to practice guideline, contribute to the decrease in morbidity and mortality rate. Bacteria, virus and fungi are the primary cause of pneumonia.8 It is referred to as Community acquired Pneumonia

Function: of DNA polymerase DNA polymerase is a very important enzyme when it comes to DNA replication. DNA polymerase, more specifically, is involved in the process of reading and adding nucleotides to the DNA strand so a complimentary stand can be made. During the DNA replication process DNA polymerase puts new nucleotides on the 3’ end of the DNA Strand. Not only does DNA polymerase add nucleotides to a DNA strand it can also act somewhat as a “proof reader”. It can pause the replication process

Vibrio fischeri is a Gram-negative bacterium that served as the model organism in this experiment. We isolated only the luxAB portion of the entire lux operon and inserted it into the pGEM vector plasmid to transcribe the luciferase enzyme required for bioluminescence. Within the lux operon, luxAB is responsible for producing subunits that form luciferase, which oxidizes the aldehyde made by luxCDE into the reduced flavin-mononucleotide FMNH2 and results in the production of light known as bioluminescence[6]

The large regulatory subunit-PriL The large subunit (PriL) of the primase is a regulatory, non-catalytic subunit, where its exact role is still undefined. Nonetheless, PriL, and in particular PriL-C-terminal domain (PriL-CTD), has been shown to have an essential role at the initiation stage of primer synthesis, but not during the extension step. In fact, it is an essential gene in yeast. It has also been inferred that PriL-CTD, is involved in the primase counting mechanism (p58 .counting..). The

2011). The vital components and techniques of gene cloning are as follows, the DNA sequence that contains the desired gene (EZH2) is amplified by Polymerase chain reaction. PCR was established by Kary Mullis in 1985, popularly known to amplify target sequences of DNA (EZH2) to a billion fold in several hours using thermophilic polymerases (Taq) ,primers and other cofactors (Sambrook and Russell, 2001). Three crucial steps are involved which are Denaturation (at 95°), Annealing of the forward