Bacterialtransformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can be transformed are called competent. Some bacteria are naturally competent (e.g B. subtilis), whereas others such as E. coli are not naturally competent. Non-competent cells can be made competent and then transformed via one of two main approaches;&n......阅读全文
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
细菌转化（bacterial－transformation）原理和操作
1．目的学会质粒DNA转化感受态受体菌的技术。2．原理质粒DNA粘附在细菌细胞表面，经过42°C短时间的热击处理，促进吸收DNA。然后在非选择培养基中培养一代，待质粒上所带的抗菌素基因表达，就可以在含抗菌素的培养基中生长。3．器材旋涡混合器，微量移液取样器，移液器吸头，1.5ml 微量离心管，双面微
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
Streaking－Bacterial－Stocks
Principle:Bacterial cells are streaked onto a medium to obtain an independent isolate. This is done to reduce the likelihood of working with a culture
Bacterial－glycerol－stocks
To 2mls of mid-log culture or 1ml of freshly saturated culture add 1 ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in l
Bacterial－cell－culture
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
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
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
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
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
In－Planta－Transformation－of－Arabidopsis
实验概要 A breakthrough in Arabidopsis research was the invention ofthe vacuum-infiltration procedure, a simple and reliable methodof obtaining
Bacterial－Media－Solutions－and－Stocks
3 agar (200 ml)Add 6 grams agar to 200 ml deionized water. Autoclave to sterilize.1.6 agar (200 ml)Add 3.2 grams agar to 200 ml deionized water.Autocl
Preparing－Overnight－Bacterial－Culture
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 1
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
L.－acidophilus－transformation
OverviewElectrotransformation procedure for Lactobacillus acidophilusProcedurePrepare Electrocompetent cellsInoculate overnight culture at 10^6 CFU/ml
Simplified－Arabidopsis－Transformation－Protocol
实验概要Our present protocol (Clough and Bent, 1998; modified from Bechtold et al. 1993) is extremely simple. We have found that the MS salts, hormone
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
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
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
Long－Term－Storage－of－Bacterial－Strains
Purpose:Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40 glycerol. It is lab policy 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
Transformation－of－E.－coli－by－Electroporation
实验概要 Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low temperature, and
Bacterial－Expression－of－GSTfusion－Proteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture (100x volume of starter culture) using the overnigh
Hydrolytic－Activity－of－Bacterial－Supernatants－for－Fungal－Suppression
As the fungal growth suppression by biocontrol agents (BCA) in solid media using dual plate assay has some issues regarding nutrient limitation. A pro
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 thr