Competentagroprepforelectroporation
day 11. Start 75 mL overnight cultures of agro (strain GV3101 C58C1 Rifr pMP90 Gmr, Koncz & Schell) in YEP in 250 mL baffle flasks.2. Grow at 28 °C, shaking.3. Prepare:~ 3.5 L sterile H2O chilled to 4 °C.10 % glycerol, autoclaved chilled to 4 °C.Centrifuge bottles (4), autoclaved.day 24. From each overnight culture, innoculate 1 L YEP in 2 L baffle flasks (1:200).5. Grow overnight at 28 °C shaking.day 36. All......閱讀全文
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
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
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
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
Plastid－Transformation－for－Abiotic－Stress－Tolerance－in－Plants
Abiotic stresses such as drought, salinity, and extreme temperatures are ?major limiting factors in plant growth and development and pose serious ?th
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
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
Chemical－transformation
Chemical transformation Preparation of chemically competent cells Have the following solutions at 0-4 deg C: a) 100 mM MgCl2 b) 100 mM CaCl2-15% gl
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
Lactobacillus－transformation
OverviewThis page details a electrotransformation protocol for?Lactobacillus?bacteria, specifically?Lactobacillus delbruckii?subsp.?bulgaricus?and?Lac
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).
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). Spi
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
Method:－Lymphocyte－Transformation
Method: Lymphocyte Transformation May 30, 1990 Rosalie Veile Principle: Lymphocytes are transformed to establish cell lines. Mononuclear
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
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
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
Biosynthesis－of－Chorismate－in－Bacteria－and－Plants
The biosynthesis of all three aromatic amino acids (tryptophan, tyrosine and phenylalanine) begins with the metabolic intermediate chorismate. The bio
Biosynthesis－of－Tryptophan－in－Bacteria－and－Plants
The aromatic amino acid tryptophan is an essential nutrient, meaning that humans and animals do not themselves have the biosynthetic machinery to synt
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
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
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
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
In－Vitro－Conservation－and－Cryopreservation－of－Ornamental－Plants
Today, the conservation of ornamental germplasm can take advantage of innovative techniques which allow preservation in vitro (slow growth storage
Verification－of－In－...
實驗概要Environmental stresses often trigger rapid generation of reactive oxygen species (ROS) in plants. Excessive amount of ROS can cause damage to
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