SimplifiedArabidopsisTransformationProtocol
(Brief version for those who are familiar with the method)Steve Clough and Andrew Bent, University of Illinois at Urbana-Champaign.Our present protocol (Clough and Bent, 1998; modified from Bechtold et al. 1993) is extremely simple. We have found that the MS salts, hormone, etc. make no difference, that OD of bacteria doesn't make much of a difference, that vacuum doesn't even make much of a differenc......閱讀全文
Simplified－Arabidopsis－Transformation－Protocol
(Brief version for those who are familiar with the method)Steve Clough and Andrew Bent, University of Illinois at Urbana-Champaign.Our present proto
Simplified－Arabidopsis－Transformation－Protocol
實驗概要Our present protocol (Clough and Bent, 1998; modified from Bechtold et al. 1993) is extremely simple. We have found that the MS salts, hormone
Transformation－Protocol－for－Arabidopsis
Transformation Protocol for Arabidopsis – AbbreviatedGerminate seed in pots↓ 4 weeksStreak bacteria onto YM/MinA↓ 2-3 days 28°CSpray/dip bacterial sus
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
In－Planta－Transformation－of－Arabidopsis
實驗概要? ? ? ? A breakthrough in Arabidopsis research was the invention ofthe vacuum-infiltration procedure, a simple and reliable methodof obtaining
Arabidopsis－RNA－extraction－protocol
1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below). Spin
Arabidopsis－RNA－extraction－protocol
1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below).Spin at 8,
Simplified－Calcium－Phosphate－Coprecipitation
Materials2x HEPES: 8 g NaCl, 0.105g NaHPO4, 6.5 g HEPES, H2O to 500 mL, pH 6.95 to 7.10 (try a range)2M CaCl2: store in aliquots at -20°CDNA 4 mg/mLPr
Arabidopsis－gDNA－isolation
This is a simple and fast protocol for the extraction of genomic DNA from Arabidopsis thaliana that works fine in PCR for simple amplicons. We only us
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
Spheroplast－Transformation
MaterialsYPD platesYPD1 M sorbitol 182 g/l (Sigma S7547)2 M sorbitol 36.4 g/100 mlSCE (per liter)1 M sorbitol (182 g)100 mM citric acid trisodium salt
Chemical－transformation
Chemical transformationPreparation of chemically competent cellsHave the following solutions at 0-4 deg C:a) 100 mM MgCl2b) 100 mM CaCl2-15% glycerolc
Lactobacillus－transformation
OverviewThis page details a electrotransformation protocol for?Lactobacillus?bacteria, specifically?Lactobacillus delbruckii?subsp.?bulgaricus?and?Lac
Cell－Suspension－Culture－of－Arabidopsis
實驗概要Cell Suspension Culture of Arabidopsis?主要試劑10% (v/v) Household BleachCallus Induction Medium?? ? ?Gamborg's B5 Basal Medium?? ? ?0.5 g/liter M
Modified－Yeast－Transformation
Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 ml saturated culture in the eve
High－Efficiency－Transformation
Day 0Make sure you have the necessary solutions (instructions for how to make them can be found here):Single-stranded carrier DNAPEG 3350 50% w/vol1.0
Agrobacterium－growth－and－transformation
Growth and storage of Agrobacterium tumefaciensStrain GV3101: resistant to gentamycin and rifampicin so add 25-50 ug/ml Gentamycin, 10 ug/ ml rifampic
Method:－Lymphocyte－Transformation
Method: Lymphocyte TransformationMay 30, 1990Rosalie VeilePrinciple:Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocyte
Fast－Yeast－Transformation
Protocol: Fast yeast transformationAdd 50 μl carrier DNA to a 1.5 ml tube.scrap cells from plate and add to the carrier DNA.Add in the following order
酵母轉化
·?????????Yeast Transformation?(Gietz Lab)LiAc/SS-DNA/PEG Transformation·?????????Yeast Transformation?(Breeden Lab)LiAc method·?????????Large-Scale Y
質粒的小量制備
·?????????Standard (alkaline lysis) Mini-Prep?(Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
質粒的小量制備
·?????????Standard (alkaline lysis) Mini-Prep?(Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
DNA轉化
DNA轉化Chemical Transformation·?????????Transformation of Competent Cells (RbCl2 Method)?(Goldberg Lab)Very nice protocol for E. Coli transformation inc
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
Production－of－Antibody－Fragments－in－Arabidopsis－Seeds
Plants offer a number of attractive benefits over conventional mammalian or bacterial cell culture systems for the production of valuable pharmace
Streptomyces:Protocols/Transformation－by－Electroporation
Description?Transform?E.coli?cells with plasmid/cosmid DNA using the method of electrophoration (inserting plasmids into?E.coli).Approx. Duration:Prep
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
L.－acidophilus－transformation
OverviewElectrotransformation procedure for?Lactobacillus acidophilusProcedurePrepare Electrocompetent cellsInoculate overnight culture at 10^6 CFU/ml
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
Transformation－of－Magnaporthe－grisea－to－phosphinothricin－resistance
Three transformation systems have been reported for the rice blast fungus?Magnaporthe grisea?(Parsons et al. 1987 Proc. Natl. Acad. Sci. USA 84:4161-4