Information on how detect and quantitate MicroRNA in laser capture microdissection samples. Includes: Obtain Laser Capture Microdissected Samples; Isolate miRNA from LCM Samples; Quantitate miRNA by qRT-PCR; Detection of miRNA in Microdissected Tissue from Mouse Brain by qRT-PCR; Differential Expression of MicroRNA in Whole Brain Tissue Compared to a More Homogeneous Population of Cells. - [Read Detect and Quantitate MicroRNA in Laser Capture Microdissection Samples]
Differences in injection of X. laevis and X. tropicalis. Includes: X. tropicalis lays eggs about 4 hours after a boost of hCG; In vitro fertilization is not nearly as efficient in trops compared to laevis; X. tropicalis embryos are much softer than X. laevis embryos; X. tropicalis embryos whose jelly coats are removed by cysteine have a loose sticky vitelline membrane; X. tropicalis do not have a "summer slump". - [Read Differences in Injection of X. laevis and X. tropicalis]
This bioassay utilises cultured H-4-II-E rat hepatoma cells to assess the aryl hydrocarbon hydroxylase (AHH) inducing potencies of planar aromatic hydrocarbons and/or contaminated environmental samples. The response of the cells to pure test chemicals or extracts of mixtures is compared with their response to the standard 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). - [Read H-4-II-E Rat Hepatoma Cell Bioassay Protocol]
Direct labeling of purified antibodies is the method of choice when simultaneously visualizing two or more antibodies of the same species, class, or subclass. This allows the localization of multiple antigens to be compared in the same cell, tissue, or sample. Labeled primary antibodies are also useful for improving background-to-readout ratios, and they can be essential for immunoassays in which good quantification is needed. - [Read Labeling Antibodies with Fluorochromes Protocol]
Protocol describes a method to determine the presence of plasmid DNA in an Agrobacterium culture. Compared to selection of transformed Agrobacterium, which can be ambiguous and normally takes several days for resistant colonies to appear, the approach described here is both rapid and accurate. - [Read PCR Analysis of Agrobacterium Protocol]
This protocol contains methods for pulling microinjection needles using two different models of pipette pullers. The advantage of pulling needles in the laboratory is that a variety of different needle types can be pulled, depending on the samples and cells being injected. An added advantage is cost; once a pipette puller has been purchased, boxes of glass capillaries are inexpensive compared to premade microinjection needles. - [Read Preparation (Pulling) of Needles for Gene Delivery by Microinjection Protocol]
To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos. - [Read Protocol Live Imaging of Caenorhabditis Elegans]
The basis of this test is that a cytotoxic chemical (regardless of site or mechanism of action) will interfere with the normal motility of the protozoan, Tetrahymena thermophila, in culture. The degree of interference of motility as compared to control cultures, related to the concentration of the test compound, provides an indication of toxicity. - [Read Tetrahymena Thermophila Ocular Irritancy Test]