GrowingOvernightCultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-capped culture tube. Large amount of culture should be made in growth flasks, using up to 25 flask volume in medium (e.g., no more than 30 mL in a 125 mL flask, or 250 mL in a 1000 mL flask).2. Touch a single colony with the end of an applicator stick and hold it in your ......阅读全文
BrdU－Labeling－Protocol
实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic
Method:－Lymphocyte－Transformation
Method: Lymphocyte TransformationMay 30, 1990Rosalie VeilePrinciple:Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocyte
Dorsal－Root－Ganglion－Neuron－Culture－from－Adult－Rats
DescriptionProcedure for culturing and maintaining Dorsal Root Ganglion (DRG) neurons from adult rats. ProcedurePreparing Culture Surface- Collagen co
Preparation－of－Bacterial－Proteins－for－Analysis－by－2DPAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term StorageMay 30, 1990Rosalie VeilePurpose:To store cell lines in a form that will insure
Master－Cell－Bank
PurposeTo describe the preparation of a Master Cell BankSafetySee SP 09-001 for lab safety considerations for the cell culture lab.EquipmentLaminar Fl
Immunocytochemistry...
实验概要The method provides a guideline procedure for staining of cell cultures using immunofluoresence.实验步骤1. General procedure 1) Coat coverslips wit
A－Method－for－Structure5
ConclusionsQuorum-sensing signaling systems involving the interaction between a signaling peptide and its cognate histidine kinase receptor are widely
Green－lab－protocol－for－vacuum－infiltration－transformation－of－Arabidopsis
This protocol is adapted from protocols by Nicole Bechtold (Bechtold et al. 1993), Andrew Bent (Bent et al. 1994) and Takashi Araki. No claims are
E.Z.N.A.®－Protocol－for－Bacteria
实验概要The E.Z.N.A. Total RNA Kit can be modified for isolation of RNA from bacterial cultures. Only cells growing at log phase should be used. Measu
Cytokine－Bioassays
Cytokine BioassaysIntroductionBiological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary cel
Biosynthetic－labeling
How long should cells be labeled? The ideal length of time to label cells depends on the protein of interest and the label that you are using. If you
Cell－Suspension－Culture－of－Arabidopsis
实验概要Cell Suspension Culture of Arabidopsis 主要试剂10% (v/v) Household BleachCallus Induction Medium Gamborg's B5 Basal Medium 0.5 g/liter M
Production－of－neuronpreferential－lentiviral－vectors
实验概要Adenoviral vectors widely used to transfer foreign genes into neuronal cells possess tropism for glial cells and are toxic to infected cells.
DNA纯化手册2
Key points to observe: a. Use a endA1- E. coli strain for plasmid propagation and isolation whenever possible. The instability of plasmids isolated fr
E.Z.N.A.™－Fastfilter－Plasmid－Mega－Protocol
实验概要The E.Z.N.A.™ family of products is an innovative system that radically simplifies extraction and purification of nucleic acids from a variety
Cosmid－DNA－Isolation
实验概要Isolation of high yields of highly pure cosmid DNA using PureLink™ HiPure Plasmid Purification Kits.实验原理The PureLink™ HiPure Plasmid Purification
Immunofluorescent－Localization－of－Tubulin
LEVEL IIMaterialsCoverslip cultures of an appropriate monolayer cell linePhosphate buffered saline (PBS)Acetone/Methanol (absolute) in a 50:50 volume
A－primary－cell－culture－model－of－rabbit－uroepithelium
Isolation of Epithelial Cells from Rabbit Bladders 1. Animal experiments were performed in accordance with the Animal Use and Care Committee. 2. Urina
慢病毒转染肝细胞方法
Lentivirus Transduction of Hematopoietic CellsMing-Jie Li and John J. RossiDivision of Molecular Biology, Beckman Research Institute of the City of Ho
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
ES－Cell－Culture－and－Manipulation2
Picking ES cell clonesOne or two days before picking colonies prepare 24-well plates of feeders. You can also use alternate protocols that utilize 96-
Isolation－and－Culture－of－Multipotent－Stem－Cells－from－Human－Bone－Morrow
Bone marrow contains three types of stem cells:1. Hematopoietic stem cells give rise to the three classes of blood cells that are found in the circu
3HThymidine－Uptake－by－Cultured－Cells
Materials Fibroblast cells in log phase growthCa , Mg free-phosphate buffered saline (PBSA)5% (w/v) Glutaraldehyde (GTA)2% (w/v) Perchloric Acid (PCA)
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Minipreparation
实验概要Plasmid DNA is isolated from small-scale (1-2 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and Solutions: Alkaline lysis
Fast－and－reliable－miniprep－RNA－extraction－from－Neurospora－crassa
We have developed a method for isolating high quality total RNA from N. crassa mycelia that reliably yields large quantities. It is possible to extrac
Multicolour－3DFISH－in－vertebrate－cells2
Small DNA-probes from cosmids or plasmids clonesThese kind of probes, especially plasmids, have become out of fashion for 3D-FISH due to their delicat
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
HThymidine－Uptake－by－Cultured－Cells
H-Thymidine Uptake by Cultured CellsLEVEL IIMaterialsFibroblast cells in log phase growthCa, Mg free-phosphate buffered saline (PBSA)5% (w/v) Glutaral