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......閱讀全文
Degenerate－PCR,－a－short－guide.
What is degenerate PCR????Degenerate PCR is in most respect identical to ordinary PCR, but with one major difference. Instead of using specific PCR pr
Degenerate－PCR
Degenerate PCR is in most respects identical to ordinary PCR, but with one major difference. Instead of using specific PCR primers with a given sequen
DNA微序列技術
·?????????Protocols for Making Drosophila Arrays?(Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,?
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
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
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
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
基因轉型(gene－transformation)
目的帶有特定基因的質體在分子生物研究上，是一個很重要的工具，將質體送入細菌的過程稱為基因轉形，經由基因轉型可使質體在細菌中大量復制，以備進一步研究。本實驗將把你在前面轉殖實驗中cDNA和質體DNA的連結反應送入細菌。 你將從本實驗學習如何進行基因轉型作用。原理早在1970年左右，有人發現細菌經由冰冷
Lactobacillus－transformation
OverviewThis page details a electrotransformation protocol for?Lactobacillus?bacteria, specifically?Lactobacillus delbruckii?subsp.?bulgaricus?and?Lac
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
ChIP－using－plant－samples－–－Arabidopsis
實驗概要This protocol describes how chromatin is prepared from Arabidopsis, which can subsequently be used for chromatin immunoprecipitation (ChIP). T
Fluorescence－In－Situ－Hybridization－using－TSA?
實驗概要This ?protocol describes steps for fluorescent in situ hybridization (FISH) ?to Drosophila embryos using Tyramide Signal Amplification (TSA?), and
Transfection－of－Mammalian－Cells－Using－Lipofectamine
Materials:???????LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium cont
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
Extraction－of－DNA－using－DNAzol?－Reagent
實驗概要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
酵母準備
Yeast DNA PreparationYeast Genomic Preparation? (Gottschling Lab)Rapid method for yeast genomic DNA isolation??Yeast DNA Preparation (rapid glass bead
人工轉錄因子的部件——人類鋅指結構2
Table 2: Binding sites and identity of ZFPs used in?VEGF?activationWe then generated artificial transcription factors by fusing the three-finger domai
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
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
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
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
Immunohistochemistry－using－AntiGanglioside－Antibodies
Immunohistochemistry using Anti-Ganglioside AntibodiesTadashi Tai~Head, Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Sc
Assay－of－Tyrosine－Kinases－Using－Synthetic－Peptides
實驗概要? ? ? ? Small synthetic peptide substrates are especially well suited for applications such as assays of tyrosine kinases in permeabilized cel
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
差異基因表達研究方法介紹（DDPCR；GENEFISHING；GENE－CHIP）
差異基因表達的研究受到了廣泛的關注，常用的技術有DD-PCR；GENE-FISHING；GENE CHIP等。簡單介紹如下： DDRT -PCR技術即mRNA差異顯示聚合酶鏈式反應技術，此技術是以PCR技術和聚丙烯凝膠電泳技術為基礎，結合銀染或放射性自顯影等顯色技術，能快速有效地