YeastGeneknockoutusingOligo／PCR
Universal primers for gene knock-out using dominant drug markers: Kan, Clonat, and Hygromisin-B. Forward primer: 5’ TCAGGGGCATGATGTGACT 3’Reverse primer: 5’ AGCTCGTTTTCGACACTGGAT 3’Plasmids for Selectable MarkersPlasmid for amplifying Kan:pKan-GenMX4.seq (GCK), product size: ~1.4kb.Plasmid for amplifying Clonat:pAG25-ClonatMX4.seq (GCK), product size: ~1.2kb.Plasmid for amplifying HB:pAG32-hphMX4.seq (GCK), prod......阅读全文
ChIP－using－plant－samples
实验概要 The immunoprecipitation (IP) of cross-linked chromatin with antibodies specific for certain histone modifications (chromatin immunoprecipitati
Adiponectin－Replenishment－Ameliorates－ObesityRelated－Hypertension（二）
Methods Animal and Animal Treatment KKAy male mice were purchased from Japan CLEA (Tokyo, Japan). This strain is a cross between black KK fema
DNA微序列技术
· Protocols for Making Drosophila Arrays (Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,
Yeast－Cell－Cycle－by－Flow－Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4
Acid－Phenol－Yeast－RNA－Prep
This is the preferred method for yeast RNA preparationuse Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fres
Use－of－the－GUS－Reporter－Gene
One of the most important considerations in the expression of heterologous proteins in plants is the choice of promoter. The study of promoter act
基因转型(gene－transformation)
目的带有特定基因的质体在分子生物研究上，是一个很重要的工具，将质体送入细菌的过程称为基因转形，经由基因转型可使质体在细菌中大量复制，以备进一步研究。本实验将把你在前面转殖实验中cDNA和质体DNA的连结反应送入细菌。 你将从本实验学习如何进行基因转型作用。原理早在1970年左右，有人发现细菌经由冰冷
Gene－Structure－Annotation－at－PlantGDB
The accurate identification of exons and introns that comprise a complete plant gene structure can be a time-consuming and challenging task. Novel
人工转录因子的部件——人类锌指结构2
Table 2: Binding sites and identity of ZFPs used in VEGF activationWe then generated artificial transcription factors by fusing the three-finger domai
Lactobacillus－transformation
OverviewThis page details a electrotransformation protocol for Lactobacillus bacteria, specifically Lactobacillus delbruckii subsp. bulgaricus and Lac
Fluorescence－In－Situ－Hybridization－using－TSA™
实验概要This protocol describes steps for fluorescent in situ hybridization (FISH) to Drosophila embryos using Tyramide Signal Amplification (TSA™), and
Method:－Cell－Counts－Using－a－Hemacytometer
Method: Cell Counts Using a HemacytometerJune 1, 1990Rosalie VeilePurpose:The purpose of this procedure is to determine the cell density of the cultur
Using－GenBank－for－Genomic－Authentication:－A－Tutorial
The GenBank database is perhaps one of the most important repositories of genetic information. A researcher working in the field of genomic authe
Extraction－of－DNA－using－DNAzol®－Reagent
实验概要DNAzol® Reagent (Genomic DNA Isolation Reagent) is a complete and ready-to-use reagent for the isolation of genomic DNA from solid and liquid sa
RNA－extraction－using－trizol/tri
RNA extraction with TRIzol (Invitrogen product name) or the equivalent TRI (Sigma-Aldrich product name) is a common method of total RNA extraction fro
ChIP－using－plant－samples－–－Arabidopsis
实验概要This protocol describes how chromatin is prepared from Arabidopsis, which can subsequently be used for chromatin immunoprecipitation (ChIP). T
Transfection－of－Mammalian－Cells－Using－Lipofectamine
Materials: LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium cont
酵母准备
Yeast DNA PreparationYeast Genomic Preparation (Gottschling Lab)Rapid method for yeast genomic DNA isolation Yeast DNA Preparation (rapid glass bead
Preparation－of－Yeast－DNA－Embedded－in－Agarose－Plugs
Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato, S. P., and A. Gnirke. 1996. RARE-cleavage analysis of YACs.
Sucrose－Density－Gradient－Fractionation－of－Yeast－Membranes
Grow a 2 ml culture, 24 hr. at 30oC in selective mediaWhen culture is ready, use it to inoculate about 55 ml (50 ml plus 5 for O.D.600 readings) of se
Reverse－Transfection－for－Gene－Function－Analysis
This guide describes a microarray-based system for the functional analysis in mammalian cells of many genes in parallel. Mammalian cells are cultured
FOSB－gene－expression－and－drug－abuse
Drug addiction is associated with long-term behavioral changes, suggesting a long-lived transcriptional regulator that responds to chronic drug exposu
Performing－a－hunt－by－interaction－mating
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt
RNAi载体pSIRENDNR－Vector
Restriction Map and Cloning Site of the RNAi-Ready pSIREN-DNR Vector. Unique restriction sites are in bold. RNAi-Ready pSIREN-DNR is provided as a lin
Immunohistochemistry－using－AntiGanglioside－Antibodies
Immunohistochemistry using Anti-Ganglioside AntibodiesTadashi Tai~Head, Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Sc
Cloning－PCR－products－using－TA－vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D. *Methods and reagents is a unique monthly column that highlights current discussions in
Sitedirected－Mutagenesis－using－PCR
Site-directed Mutagenesis using PCRMichael P. Weiner, Tim Gackstetter, Gina L. Costa, John C. Bauer, and Keith A. KretzFrom: Molecular Biology: Curren
Detection－of－apoptotic－process－in－situ－using－immunocytochemical
1. INTRODUCTION Apoptosis was observed from invertebrates to lower and higher verterbrates, and intervenes both in physiological and in pathological
Assay－of－Tyrosine－Kinases－Using－Synthetic－Peptides
实验概要 Small synthetic peptide substrates are especially well suited for applications such as assays of tyrosine kinases in permeabilized cel
FACS－Analysis－Using－Peripheral－Blood－Cells
FACS Analysis Using Peripheral Blood CellsCollect blood (75 microliters) into 1ml PBS containing 5 microM EDTA (10 microliters of 0.5 M stock) and mix