All detection assays were performed on a standard 96 well test plate. The 96 sample test plate was comprised of genomic DNA of representative mosquito individuals of all the known kdr genotypes including three individuals heterozygous for both the east and west kdr alleles. The plate included DNA from An. gambiae s.s (both S and M forms) An. arabiensis, Anopheles quadriannulatus, Anopheles melas, Anopheles merus and Anopheles funestus. The amount of DNA was variable between samples to test the sensitivity of each assay. DNA concentration was determined by absorption at 260 nm using a NanoDrop spectrophotometer (NanoDrop Technologies). The plate also included a number of Plasmodium falciparum DNA samples and water blanks as negative controls. The details of each of the 96 samples (including species, molecular form, collection location, DNA concentration and kdr genotype) is given in Additional file
MELAS Syndrome
This progressive condition can lead to a range of neurological and muscular symptoms, including seizures, dementia, and muscle weakness.
Through PubCompare.ai's AI-driven protocol optimization, researchers can enhance the accuracy of their MELAS studies by quickly identifying the most effective research protocols from literature, preprints, and patents.
This intelligent comparison platform helps optimize the research process and deliver deeper insights into this complex disorder.
Most cited protocols related to «MELAS Syndrome»
All detection assays were performed on a standard 96 well test plate. The 96 sample test plate was comprised of genomic DNA of representative mosquito individuals of all the known kdr genotypes including three individuals heterozygous for both the east and west kdr alleles. The plate included DNA from An. gambiae s.s (both S and M forms) An. arabiensis, Anopheles quadriannulatus, Anopheles melas, Anopheles merus and Anopheles funestus. The amount of DNA was variable between samples to test the sensitivity of each assay. DNA concentration was determined by absorption at 260 nm using a NanoDrop spectrophotometer (NanoDrop Technologies). The plate also included a number of Plasmodium falciparum DNA samples and water blanks as negative controls. The details of each of the 96 samples (including species, molecular form, collection location, DNA concentration and kdr genotype) is given in Additional file
A cross between females of the GA-CAM (a M-form colony originated from field gravid females collected in Cameroon) and males of the GA-BF-5.7 colonies (a S-form colony originated from a single field gravid female collected in Burkina Faso) was performed and parental individuals and F1 hybrid females were analysed.
The blind species identification trials were performed using five 96 well test plates containing 466 samples. Samples were field collected from Cameroon, Ghana, Kenya, South Africa, Malawi, Sao Tome, La Reunion, Tanzania, Sudan, Angola, Burkina Faso, Gabon, and Mozambique. These samples had been initially identified to species at the time of collection using the standard PCR method and included 169 An. gambiae s.s., 173 An. arabiensis, 66 An. quadriannulatus, and 21 samples of An. melas and An. merus, the remaining samples were either undetermined or negative controls. This information was withheld from the persons who carried out the testing of each assay to ensure no bias occurred in the scoring of results. For all samples DNA was extracted from single mosquitoes using either the Livak or Ballinger Crabtree methods [19 (link),20 (link)]; DNAzol reagent (Molecular Research Centre, Inc) at one-fifth the recommended reagent volume for each extraction or using a crude boil and centrifugation protocol. This was a variant of the STE method of O'Neill et al. [21 (link)] in which samples were ground in STE buffer (100 mM NaCl, 10 mM Tris-HCl, pH8.0, 1 mM EDTA, pH8.0), heated to 95°C for 5 mins, centrifuged for 3 mins at 13,000 rpm and the supernatant used directly as PCR template. The DNAs were resuspended in either TE buffer or sterile water at volumes between 100 and 200 μl. To determine the sensitivity of the three identification methods a dilution series of the DNA from each of the five main species in the Anopheles gambiae complex was included in the trial. For this, DNA preparations were diluted to 20 ng/ul (as determined by absorption at 260 nm using a NanoDrop spectrophotometer, NanoDrop Technologies). The samples were then serially diluted down to a 1 in 1 × 06 dilution.
Most recents protocols related to «MELAS Syndrome»
Example 7
The purpose of this experiment was to visualize engineered meganuclease localization when the nuclear localization signal (NLS) on the protein was replaced with a mitochondrial transit peptide (MTP).
8e5 MELAS cybrid cells were nucleofected with 1e6 RNA copies/cell engineered meganuclease mRNA using the Lonza 4D-Nucleofector™ (SF buffer, condition CA-137). Two engineered meganuclease constructs were compared: one without a targeting sequence and one with an MTP at the N-terminus of the protein. At 24 hours post-nucleofection the cells were stained with 50 nM MitoTracker™ Deep Red FM (ThermoFisher Scientific, M22426) and Hoechst 33342 (1:5000 dilution) for 30 minutes Cells were imaged using the Zeiss microscope using 63×Z-stack images.
Without a targeting sequence, engineered meganuclease staining appears diffuse throughout the cytoplasm and the nucleus (
With the addition of an N-terminus MTP, engineered meganucleases are effectively localized to the mitochondria.
The study sites in southern Ghana
Dodowa (5° 52′ 58.3212ʺ N 0° 5′ 52.9548ʺ W) is a community located in the savannah-forest transition zone in the Shai Osudoku District and is about 39 km from the city of Accra. It has an average temperature of 27 ℃ with a bimodal rainfall pattern like Anyakpor. It has secondary forest type vegetation with little original virgin forest left as a result of deforestation. Due to intense human activities in Dodowa, water accumulates at construction sites, unpaved roads and low-lying areas creating suitable larval habitats for mosquitoes.
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More about "MELAS Syndrome"
This progressive condition is characterized by Mitochondrial Encephalomyopathy, Lactic Acidosis, and Stroke-like Episodes, leading to a range of debilitating symptoms.
Researchers studying MELAS Syndrome often face challenges in optimizing their research protocols to ensure accurate and insightful findings.
However, with the help of AI-driven platforms like PubCompare.ai, scientists can now quickly identify the most effective research protocols from literature, preprints, and patents.
By utilizing this intelligent comparison platform, researchers can enhance the accuracy of their MELAS studies and gain deeper insights into this intricate disorder.
The platform helps optimize the research process by highlighting the most effective protocols, which may incorporate techniques like DMEM (Dulbecco's Modified Eagle Medium), FBS (Fetal Bovine Serum), QIAxtractor robot, DNeasy Blood and Tissue Kit, StemFit AK02N medium, DMEM/F12, Uridine, Apal1, Y-27632, and SB203580.
Through this AI-driven approach, researchers can unlock new understanding of the underlying mechanisms of MELAS Syndrome, leading to improved diagnosis, management, and treatment strategies for patients suffering from this debilitating condition.
OtherTerms: Mitochondrial disorder, Mitochondrial Encephalomyopathy, Lactic Acidosis, Stroke-like Episodes, MELAS, Neurological symptoms, Muscular symptoms, Seizures, Dementia, Muscle weakness, Research protocols, Protocol optimization, PubCompare.ai, DMEM, FBS, QIAxtractor robot, DNeasy Blood and Tissue Kit, StemFit AK02N medium, DMEM/F12, Uridine, Apal1, Y-27632, SB203580.