SliceandExplantCultureProtocols–Hevnerlab2002

for axon tracing & cell culture studies in vitro using embyros age E11.5 – E16.5modified from Rubenstein lab and Price lab protocols1. Setupa. 2 hr before dissections:1. Prepare 1X Krebs (1000 ml) and cool on ice. 2. Prepare 100 ml of sterile filtered Krebs buffer and cool on ice. 3. Prepare 100 ml serum–supplemented medium, and place in incubator set at 37°C and 5% CO2. 4. Prepare 100 ml serum–fre......阅读全文

Slice-and-Explant-Culture-Protocols-–-Hevner-lab-2002

for axon tracing & cell culture studies in vitro using embyros age E11.5 – E16.5modified from Rubenstein lab and Price lab protocols1. Setupa. 2 hr be

Human-Embryonic-Stem-(ES)-Cell-Protocols——General-notes-on-ES-cell-culture

hES media has a two week shelf life.hES cells should be cultured in 4, 6, 24, 48, 96 well plates. Growing cells in flasks is not recommended because i

显微镜技术——电子显微技术

The Transmission Electron Microscope (TEM) (HEI)An explanation of how the TEM works.  TEM Specimen Preparation (HEI)  Serial Sectioning (Walter Steffe

CDNA文库

 CDNA文库(主要内容如下)·         Construction of cDNA Library·         Construction of Genome DNA Library·         Library Screening  OthersConstruction of cD

流式细胞仪技术专辑

Flow Cytometry Analysis (Springer Lab, Harvard University) Flow cytometry employs instrumentation that scans single cells flowing past excitation sour

流式细胞仪技术专辑

 最方便的实验干货查询工具微信扫码进入「丁香实验」小程序编辑: 呜咽分享到:      Flow Cytometry Analysis (Springer Lab, Harvard University)Flow cytometry employs instrumentation that scan

粘粒

Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re

粘粒

Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re

M13噬菌体

·         M13 Phage (Michael Blaber)Very useful background information about M13: its infection, replication, packing, cloning. If you are new to phag

LCM-PROTOCOLS

Slide SectioningParaffin blocks- For DNA analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap™, rinsed thoro

ORGANOTYPIC-LIMB-CULTURES

-embryos are dissected from timed-pregnant mice from 10.5 - 11.5 d.p.c.-limb buds are microdissected and placed in holding medium (L15 medium suppleme

ORGANOTYPIC-KIDNEY-CULTURES

-embryos are dissected from timed-pregnant mice from 11.5 d.p.c. to 13.5 d.p.c.-metanephroi and associated ureteric buds are microdissected and placed

The-OP9DL1-System:-Generation-of-TLymphocytes-from-Embryonic1

The OP9-DL1 System: Generation of T-Lymphocytes from Embryonic or Hematopoietic Stem Cells In VitroRoxanne Holmes and Juan Carlos Zúñiga-Pflücker1Sunn

Bacteria-Growth-and-Culture-Bacteria-Growth-and-Culture

Flagella and motilitymonotrichous flagella - the bacterial cell has a single flagellaperitrichous flagella - the bacterial cell has several flagella w

Streptomyces:Protocols/Conjugation

Intergeneric Conjugation and OverlayDescription Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient str

Neutralizing-Bioassay-Protocols

Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri

DAPI-Counterstaining-Protocols

实验概要The  blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA;  it appears to associate with AT clusters in the minor groove. Binding

Smolke:Protocols/Western

OverviewBlotting for large V5-tagged proteins in S. cerevisiaeMaterialsY-PER (Pierce)Halt EDTA-free Protease Inhibitor (Pierce)NuPAGE Novex Bis-Tris 4

CGH-Protocols-(二)

DNA preparation by cryotom tissue dissectionPreparations/Materials: Cool cryostat down to -20 to -30°C about 3 hours prior to dissection Label eppendo

General-Cloning-Protocols

Large Scale Preps: (See Large scale plsasmid prep protocol for more details)Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a.m. w

Western-Blotting-Protocols

back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.

Streptomyces:Protocols/PCR

Description Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template doubl

CGH-Protocols-(三)

Hybridizationreagents: labeled tumor and normal-DNA (see protocol Nick translation) salmon sperm DNA, 10 mg/ml (e.g. Promega) human Cot1 DNA, 1 mg/ml

CGH-Protocols-(一)

Metaphase chromosome preparationMaterials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best

CGH-Protocols-(四)

CGH Image acquisitionImages were acquired through a Zeiss Axiophot fluorescence microscope using a Plan NEOFLUAR oil objective x63, N.A. 1.25 (Zeiss,

Sectioning-stained-embryos.

The protocols for plastic and wax sections as used by the Vize lab. These protocols work, but they have not been optimized. If anyone has better proto

Lactobacillus-culture

OverviewGeneral overview and guidelines on how to grow up a culture of LactobacillusMaterialsMRS broth (difco)MRS agar (difco)anaerobic conditionsProc

The-OP9DL1-System:-Generation-of-TLymphocytes-from-Embryonic4

TROUBLESHOOTINGProblem: The OP9 cells are more than 80%-90% confluent.Solution: It is important when creating working stocks of OP9 cells for freezing

Streptomyces:Protocols/Spore-Prep

Spore Prep - Inoculating & HarvestingDescription A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 2

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