Preparationofcytospinfromsinglecellsuspension.

Cell number, speed and trivial details affect cytospin .Basic protocol:Prepare a cell suspension of not more than 0.5 x 106 cells / ml of protein-containing medium. Pre-label the slides.Procedure for old type cytocentrifuge (be careful, this is not biohazard proof!!)Prepare the slides mounted with the paper pad and the cuvette in the metal holder . Load up to 200 µl of this suspension in each cuvette.&......阅读全文

酵母转化

·         Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation·         Yeast Transformation (Breeden Lab)LiAc method·         Large-Scale Y

RNA提取

RNA提取(主要内容如下)Tips for Handing RNA Total RNA IsolationmRNA IsolationrRNA IsolationOthersQ & A posted in the Method Forum  Basic Procedures for Handing

单细胞测序技术(single-cell-sequencing)-综述(二)

DroNC-seq10月,Broad研究所张锋教授团队在Nature Methods上发表题为“Massively Parallel Single-Nucleus RNA-Seq with DroNc-Seq (doi: 10.1038/nmeth.4407)” 的文章,提出了一个新的测

单细胞测序技术(single-cell-sequencing)-综述(一)

单细胞生物学最近几年是非常热门的研究方向。在这一领域中,最前沿的则是单细胞测序技术。传统测序方法一次处理成千上万个细胞,得到的变异水平也是成千上万个细胞的平均后水平。但是,就如同世界上没有完全相同的两片树叶一样,没有两个细胞是完全相同的。所以,单细胞测序对于研究单个细胞就显得至关重要。单细胞测序可以

自动化的微流控芯片系统在单细胞中检测MicroRNA的异质性3

结论·  我们在C1TM单细胞自动制备系统开发了一种简洁的实验方案,能以最少的手工操作,在不到24小时内,平行处理高达96个单细胞,对其miRNA表达谱进行分析。·  C1 miRNA STA实验方案使用了Life Technologies为miRNA优化过的试剂。特别的,Megaplex™ RT及

Combined-Flow-Cytometric-Measurement-of-Two-CellSurface-Antigens

INTRODUCTIONFlow cytometry is frequently used to assess nucleic acid content in individual cells. Based on DNA content alone, however, cells in the qu

DNA抽提

DNA抽提(主要内容如下)·   Working with DNA·   DNA Extraction from Bacteria and Other Organisms·   DNA Extraction from Cell and Tissue·   Mitochondria DNA Isola

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

Isolation-of-Cell-Wall-Proteins-from-Medicago-sativa-Stems

Plant cell walls are highly dynamic and chemically active components of plant cells. Cell walls consist primarily of polysaccharides, with protein

Revised-protocol-for-establishing-a-mammary-cell-line-from-tumors

Revised protocol for establishing a mammary cell line from tumorsDrench mouse in 70% ethanol, pin and dissect openSterile dissection of tumor- leave s

Method:-Lymphoblastoid-Cell-Lines-from-Frozen-Whole-Blood

Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31, 1990Rosalie VeilePurpose:Blood Samples can be stored frozen as a backup in case an LC

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

噬菌体的生长

Preparing Lawn Cells for M13 Cloning (Life Technologies)Lawn cells require the F' episome for M13 infection and may be prepared  Streaking Lambda

体外荧光法检测核内体早期动力学

A fluorescence-based in vitro assay for investigating early endosome dynamicsSina V Barysch1,2, Reinhard Jahn1 & Silvio O Rizzoli2ABSTRACTEarly endoso

Detection-of-apoptotic-process-in-situ-using-immunocytochemical2

B) TUNEL in situ procedureB.2.1 Materialsproteinase K (pK) (A2), H2O2 , TdT buffer (A1), TdT enzyme (A2), biotinylated dUTP (A2), TB buffer (A1), seru

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

噬粒(phagemid)

Phagemid Related Protocol (Erik)provides procedures for phagemid lysate preparation, titering phagemid, determining uracil incorperation and SS DNA pr

Mitochondrial-DNA-Isolation-from-Somatic-Embryogenic-Cell-Cultures-of-Larix

Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) and Radetzky (1990). Cell cultures at four days

ASENSITIVE-METHOD-FOR-DETECTION-OF-APOPTOSIS-BY-SINGLE-LASER-FLOW-CYTOMETRY

MATERIALS:1. 1 X PBS (PBSAz, 1 X PBS, e.g., Irvine Scientific, CA, containing 2% newborn calf serum and 0.1% sodium azide)2. 7-Amino-actinomycin D (7-

质粒的小量制备

·         Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip

质粒的小量制备

·         Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip

Lambda噬菌体

·         Lambda DNA Preparation (Stanford DNA Sequence & Technology Center)Detailed protocol for lambda DNA preparation with recipes·         Isolati

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

Chemotaxis-Assay-Using-Microchemotaxis-Chambers-(Modified-Boyden-Chamber)

Overview This assay uses a simple apparatus to study chemotaxis of leukocytes or other migratory cells. The apparatus consists of two multi-well chamb

蛋白质提取和纯化

蛋白质提取和纯化(主要内容如下)Protein Extraction Protein PurificationProtein PrecipitationColumn PreparatioinQ & A Posted in the Method ForumProtein ExtractionWhole

An-Integrative-Procedure-for-Apoptosis-Identification-and-Measurement2

TroubleshootingCritical Steps(1) Don’t trypsinize cells for too long when collecting them.(2) Rotation speed should be no more than 1500 rpm during ce

Placental-trophoblast-and-chorionic-cell-cultures

1. Placental trophoblast and chorionic trophoblast cells were prepared using a modification of the method. 2. Term human placentae and chorion tissue

酵母准备

Yeast DNA PreparationYeast Genomic Preparation  (Gottschling Lab)Rapid method for yeast genomic DNA isolation  Yeast DNA Preparation (rapid glass bead

组织学——显微解剖

Laser Capture Microdissection (LCM)Introduction to LCM  (BJMU)  Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections (NIH Laser Capture M

Antibody-Purification-using-Protein-A,-Protein-G,-or-Protein-L-Agarose

实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants. It should