Agarose gel electrophoresis

A colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT, Sigma-Aldrich Chemical Co.) assay was performed to measure the cell viability. Briefly, RAW 264.7 cells were treated with different concentrations of spermidine for 24 h or pretreated with spermidine for 1 h before stimulation with LPS for 24 h. After incubation, the medium was discarded, and MTT solution (5 mg/mL in phosphate-buffered saline, PBS) was added to each well and incubated for another 3 h at 37 °C. The…

then analyzed in a process called agarose gel electrophoresis. Because DNA contains phosphate groups, which are negatively charged, applying an electrical current across the DNA will cause it to move towards the negative side of the gel. Since the restriction enzymes have separated the DNA into varying lengths, the rate at which the DNA fragments move through the gel will be dependent on their size. The smaller fragments will be able to more rapidly traverse the gel while the larger fragments…

lab’s interests have clinical implications through our work with macrophage migration inhibitory factor, a pro-inflammatory cytokine molecule associated with rheumatoid arthritis. So far, I have been able to help set up experiments, run agarose gel electrophoresis, determine protein and DNA concentration, and perform a bacterial transformation experiment. These experiences have been very satisfying, as I am working with and manipulating the things I read about in my textbooks. I look forward to…

The Blooding The Blooding, written in 1989 by Joseph Wambaugh, relates the story of a two English girls brutally raped and strangled three years apart in the 1980’s. The novel follows the investigation of the Narborough murder and how the discovery of a new forensic technique was vital to solving the case and finding the killer. This discovery of genetic fingerprinting by Alec Jeffreys during the time of this investigation revolutionized the world of forensic science. The novel begins by…

Chitin is the second most abundant natural polysaccharide consisting of β(1→ 4)-linked N-acetyl-D-glucosamine (GlcNAc) units in a linear form. Chitin is insoluble in water and mainly exists in two crystalline (α- and β-) forms. The α-chitin consists of sheets of tightly packed alternating parallel and antiparallel chains (1) and is abundant in the exoskeleton of arthropods, insects, fungi, and yeast cell walls. The chains are arranged in parallel orientation in β-chitin (2), which occurs less…

2.5. Morphological Identification/Evaluations of Entomopathogenic Fungi The six isolates of fungal entomopathogens were provided by Department of Entomology, Penn State USA. Preliminary morphological identifications of fungal entomopathogens such as color of colony, mycelial characteristics and spores from all six isolates were done at microscopic level. 2.6. Molecular Characterizations of Entomopathogenic Fungi Cultures of Metarhizium fungus were grown in nutrient broth (Bacto, Australia) in…

Saibhuvaneswari RA1611014010048 JUMPING GENES Transposable elements (TEs), also known as "jumping genes" or transposons, are sequences of DNA that move (or jump) from one location in the genome to another. A transposable element (TE or transposon) is a DNA sequence that can change its position within a genome, sometimes creating or reversing mutations and altering the cell's genetic identity and genome size. Transposition often results in duplication of the TE. The transposable elements make…

Laboratory Diagnosis Monitoring Parameters • Tourniquet Test Acts as an emergency diagnostic tool whereby formation of more than 20 petechiae per square inch indicates a positive tourniquet test. A circle of 2.5 cm in diameter may be drawn and the number of petechiae in it is counted. More than five petechiae indicates positive test. • Complete Blood Cell Count Blood tests present with lowered total white blood cell, platelet and neutrophil counts and a 20% or more increase in haematocrit…

uses the deoxynucleotides (dNTPs) to create the 16s rRNA copy. After repetitive cycles of denaturation, annealing, and extension through optimal temperatures, copies of the 16s rRNA band were made and purified to later be assessed using agarose gel electrophoresis. Based on Figure 7, there was a clear indication of a band on 16s rRNA PCR with a basepair length of approximately 1450bp. By analyzing this band in terms of its specific nucleotide sequence and comparing it with the national…

the solution from the previous experiment is needed, varying tubes, master mix, ice, and a PCR machine. For experiment 7 varying tubes are needed, the product from the previous lab, pipette, ice, heat block, SOC, and incubator. For experiment 8 a agarose gel is needed along with ice, pipette, toothpick, centrifuge, vortex, buffer P1, buffer P2, buffer N3, buffer PE, and sterile water. For experiment 9 a NanoDrop machine is needed, a pipette, buffer, and incubator. Lastly for experiment 10 the…