TransformationofElectrocompetentE.coliwithBlue／Whiteselection
Desalt DNA template by EtOH precipitation in NaOAc followed by at least 2x washes with 70% EtOH. Resuspend in 5 - 15 µL of sterile H2O.Rinse cuvettes (if they have been used before) 5x with deionied H2O, and place them on ice. This is sufficient to avoid background growth in most cases.Set a BioRad MicroPulser to "Ec1" for 1 mm cuvettes, or "Ec 2" for 2 mm cuvettes.Electroporate the DN......阅读全文
Transformation－of－Electrocompetent－E.－coli－with－Blue/White－selection
Desalt DNA template by EtOH precipitation in NaOAc followed by at least 2x washes with 70% EtOH. Resuspend in 5 - 15 µL of sterile H2O.Rinse cuvettes
Antibiotic－concentrations－for－E.－coli－selection
Antibiotic ConcentrationsThe following list shows recommended antibiotic concentrations for LB media or agar plates.AntibioticConcentrationAmpicillin1
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
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
Selection－of－Transfected－Suspension－Cells
Contributor: Suprya JayadevDate: December 13, 19941) Transfect cells.2) Culture cells 1-3 passages in a T-75 flask containing selection material (e.g.
DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
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
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
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
Agar－Plates－for－Selection－of－Clones－in－Bacteria
Cloning of PCR productsStocks:LB Agar: Luria Broth after Lennox:per LiterTryptone10 gYeast Extract5 gSodium Chloride5 gBact. Agar15 g pH 7.0Autoclave
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
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
Method:－Lymphocyte－Transformation
Method: Lymphocyte TransformationMay 30, 1990Rosalie VeilePrinciple:Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocyte
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
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
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
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
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
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
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
Inoue法制备大肠杆菌超级感受态细胞
实验步骤： 1、Inoculate from an overnight grown in LB.从培养过夜的LB平板上挑取单菌落 。2、Grow in 250 ml "SOB" at 18℃ until OD600 = 0.6.(0.3)接种于250ml SOB，18度培养至OD＝0.6。3、On
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
AgrobacteriumMediated－Maize－Transformation:－Immature－Embryos－Versus－Callus
Transformation with Agrobacterium tumefaciens is the preferred method for delivery of transgenes into a wide range of plant species including maiz
Phospholipid－Biosynthesis－in－E.－Coli－Pathway
The biosynthesis of membrane phospholipids occurs through distinct pathways in mammals and bacteria. In the mammalian pathway for the synthesis of pho
Maintenance－of－Probes－in－bacteria－including－Escherichia－coli
Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on
Cell－cycle－analysis－of－Escherichia－coli－cells
Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of a round o