SleepingBeautytransposonmutagenesisinratspermatogonialstemcells
Sleeping Beauty transposon mutagenesis in rat spermatogonial stem cellsZoltán Ivics,1, 2 Zsuzsanna Izsvák,1, 2 Gerardo Medrano,3, 4 Karen M Chapman3, 4 & F Kent Hamra3, 4Nature Protocols Volume: 6, Pages: 1521–1535 Year published: (2011) DOI: doi:10.1038/nprot.2011.378 Published online 08 September 2011 AbstractWe describe an experimental approach for generating mutant alleles in rat spermatogonial stem......阅读全文
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Gene-splicing-and-mutagenesis-by-PCRdriven-overlap-extension
实验概要 Extension of overlapping gene segments by PCR is a simple, versatile technique for site-directed mutagenesis and gene splicing.Initial
Rat-Blood-Collection-Protocols
实验概要The procedure presented below describes a method for collecting rat blood.实验步骤Rat should be fully anesthetized (e.g., unresponsive to toe pinch).1
Differentiate-ES-cells-into-glial-cells-and-neurons
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.___________________Day 1: Trypsiniz
Culture-of-BEND-Cells-(Bovine-Endometrial-Cells)
Culture of BEND Cells (Bovine Endometrial Cells)Charles E. Krininger, III and Peter J. Hansen Dept. of Animal Sciences, University of FloridaThis prot
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RAT/MOUSE-GROWTH-HO...
实验概要This Rat/Mouse Growth Hormone ELISA kit is used for the non-radioactive quantification of Growth Hormone in rat or mouse serum, plasma, tissue
Preparing-Mitochondria-from-Rat-Liver
Liver is a convenient source for functional intact mitochondria for a number of reasons. Animal tissue is more readily homogenized than plant tissue b
Isolation-of-rat-cardiac-fibroblasts-and-cardiomyocytes
1. Hearts were removed from newborn rats (day 0), put into calcium- and bicarbonate-free HEPES-buffered Hanks’ medium, cut into pieces and digeste
Preparation-of-Rat-Liver-Cell-Cytosol
These protocols should yield enough cytosol and organelles for 1-200 MT/Organelle motility assays.Solutions and Reagents Freshly removed or flash fro
Preparation-of-Mitochondria-from-Rat-Liver
Preparation of Mitochondria from Rat LiverRat liver is an ideal source for functional intact mitochondria for a number of reasons. We use Sprague-Dawl
Histopathological-Approach-to-Rat-Liver-Tissue
ProcedureAfter deep ether anesthesia, dissect the rat’s liver (Wistar albino rats, 200 – 250 g) by cutting on the ventral side.Fix 2 – 3 mm. of the li
Immunofluorescent-Staining-of-Mouse-and-Rat-Leukocytes
I. ProcedureHarvest cells from tissue, preparing a single cell suspension. Red blood cells may be removed by lysis or density gradient: Red blood cell
mice-islet-isolation
1. Islets of Langerhans were isolated from 5- to 7-week-old nonobese diabetic (NOD) mice. 2. Which involves cannulation of the common bile duct and
Lyophilizing-Cells
Lyophilizing CellsInoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized.Incubate the culture at 37°C
Trypsinizing-cells
There are many procedures with which to trypsinize cells. All include washing the cell monolayer with TD, or in rare cases, with VE. This removes seru
Lyophilizing-Cells
Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized.Incubate the culture at 37°C with vigorous shak
Growing-cells
No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irrespe
Freezing-Cells
1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g.
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Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
Preparing-cells-and...
实验概要The method provides a protocol and tips for BrdU staining in tissue sections.Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic