PreparationandStainingofFrozenTissueSections
I. Preparation of Frozen Sections for SectioningMaterials neeed:2-methylbutane (isopentane)Liquid NitrogenDry icePeel-Away?/sup> base moldsFrozen tissue matrix (OCT?/sup> or Cryomatrix?/sup>)Long forcepsNecropsy toolsSuperfrost Plus slidesLabel base mold and partially fill the mold with frozen tissue matrix.Sacrifice animal by prescribed and approved euthanasia techniques.Remove desired tissues, trim and cut......阅读全文
CELL－MEMBRANE－PREPARATION
I. Solutions: A. Ca and Mg free Phosphate Buffered Saline (PBS) solution, buffered with 0.02M Hepes. pH=7.4 B. Ca and Mg free PBS, buffered with
Preparation－of－human－platelets
Preparation of human platelets 1. Human blood was taken from drug-free volunteers on the day of the experiment using acidic citrate dextrose
HELPER－PHAGE－PREPARATION
HELPER PHAGE PREPARATION1. Grow an overnight of NM522 in NZCYM medium.2. Dilute overnight 1:100 and grow to an A600 = 0.3 (@2.5 x 108 cells/ml).3. Inf
Preparation－of－Polyacrylamide－Gels
1. Prepare 20X TBE as:216 g Tris Base110 g Boric Acid80 mL 500 mM EDTA, pH 8.0700 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Prepare Acrylamide so
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
Plasma－and－Serum－Preparation
实验概要Serum is the liquid fraction of whole blood that is collected after the blood is allowed to clot. The clot is removed by centrifugation and the
Preparation－of－Mouse－Neutrophils
实验概要Preparation of Mouse Neutrophils实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline so
Competent－Cell－Preparation
实验概要Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. These cells readily incorporate f
Preparation－of－Agar－plates
Prepare media and add 1.5 agar before autoclaving it (15g per liter). After autoclavation, cool the media in a 55 degree waterbath. Do not allow the s
Metaphase－chromosome－preparation
Materials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best.-Nr. 295892) cell cuture flask
Preparation－of－Phage－Lysates
Preparation of Phage LysatesInoculate 5 ml of lambda-broth in a glass culture tube with a single colony of an appropriate host strain of E. coli. Incu
DGK－Membrane－Preparation
Reagents:Bacterial strainE. coli N4830/pJW10LB amp media50 µg/ml ampicillinHigh salt bufferfor 1 L50 mM KH2PO4 6.8 g150 mM KCl 11.18 g50 mM sodium pyr
Preparation－of－Mouse－Neutrophils
实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
PREPARATION－OF－SEQUENCING－GELS
MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc.
PREPARATION－OF－MICROINJECTION－PIPETTES
INJECTION AND HOLDING PIPETTESThe glass capillary tubing used should be thin walled, borosilicate glass without a fibre.e.g. Clark Electromedical Inst
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
实验概要RNA analysis on non-denaturing agarose gel electrophoresis实验步骤1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer instead of 1X TBE- use agarose gel in the concentration of 1.1%-1.
Isolation－and－growth－of－mouse－primary－myoblasts
Isolation of limb muscle from neonatal mice1. Neonatal mice by decapitation or CO2 inhalation.2. Rinse the limbs with 70% ethanol and remove them wi
Embryonic－limb－bud－culture－in－media
Early in embryonic development, the region of the chick embryo which is determined to form a limb first differentiates from the rest of the embryo1. T
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
Cell－Surface－Immunofluorescence－Staining－Protocol
实验概要A method of identifying and enumerating specific cell types in a heterogeneous population of cells by enhancing the specific staining of desired
Immunofluorescent－staining－of－Sea－Urchin－embryos
1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 1
SSR－GEL－and－Silver－Staining－Protocol
I. EQUIPMENT:DNA sequencing unit (35 x 45 cm) & 2000V power supplyClampsLg. plastic trays (4), about 43 x 50 x 8 cm, and one lidTwo rocking platformsH
Methylene－Blue－DNA－staining－protocol
Methylene Blue DNA staining protocolProtocol:Load 2-5X the amount of DNA that would give bands of moderate intensity on an ethidium bromide stained ge
Intracellular－Immunofluorescent－Staining－for－Flow－Cytometry
IntroductionA modification of the basic immunofluorescent staining and flow cytometric analysis protocol can be used for the simultaneous analysis of
组织学——试剂和溶液
Fixative Preparation10% Formal Saline Preparation (Nottingham Pathology Lab) 4% Paraformaldehyde Preparation (Nottingham Pathology Lab) Fixatives (XM
Sample－preparation－(analytical－gels)
Sample preparation and solubilization are crucial factors for the overall performance of the 2-D PAGE technique. Protein complexes and aggregates shou
Cryopreservation－and－Recovery－of－Mature－Differentiated－Neural－Cells
实验概要Primary neuronal cultures are indispensable in the field of neurobiology and pharmacology. Many researchers favor freshly isolated neuronal cell
Tissue－Culture－Methods3
REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach student should m
Synthesis－and－Probing－of－Membranebound－Peptide－Arrays1
Synthesis and Probing of Membrane-bound Peptide ArraysRonald FrankDepartment of Chemical Biology, GBF (German Research Center for Biotechnology), 3812