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......閱讀全文
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
細菌轉化（bacterial－transformation）原理和操作
1．目的學會質粒DNA轉化感受態受體菌的技術。2．原理質粒DNA粘附在細菌細胞表面，經過42°C短時間的熱擊處理，促進吸收DNA。然后在非選擇培養基中培養一代，待質粒上所帶的抗菌素基因表達，就可以在含抗菌素的培養基中生長。3．器材旋渦混合器，微量移液取樣器，移液器吸頭，1.5ml 微量離心管，雙面微
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－Colony－PCR
Bacterial Colony?PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
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
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
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
Method:－Lymphocyte－Transformation
Method: Lymphocyte TransformationMay 30, 1990Rosalie VeilePrinciple:Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocyte
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
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
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
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
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
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
L.－acidophilus－transformation
OverviewElectrotransformation procedure for?Lactobacillus acidophilusProcedurePrepare Electrocompetent cellsInoculate overnight culture at 10^6 CFU/ml
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
DNA轉化
DNA轉化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－E.－coli－by－Electroporation
實驗概要? ? ? ? Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low?temperature, and
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
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
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
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