MaxiprepofplasmidDNAfromE.coli
IngredientsIngredients are per culture; make enough for one extra culture to allow for pipetting error).150μL sterile 50% glycerol1mL TEG (25mM Tris-Cl, 10mM EDTA, 50mM dextrose)111μL 20mg/mL lysozyme2mL worth of components for solution 2: 200μL 10% SDS, 100μL 4M NaOH, 1.7 mL autoclaved water1.5mL Solution 3: 3M K+, 5M acetate (3M potassium-acetate, 2M acetic acid -- glacial is 17M)3.7mL isopropanol1mL TE buffer0.5mL......阅读全文
Maxiprep－of－plasmid－DNA－from－E.－coli
IngredientsIngredients are per culture; make enough for one extra culture to allow for pipetting error).150μL sterile 50% glycerol1mL TEG (25mM Tris-C
Maxiprep－of－plasmid－DNA－from－E.coli－protocol
Solutions/reagents:LB broth + selective marker50% sterile glycerolTEG(25mM Tris-Cl, 10mM EDTA, 50mM dextrose)20 mg/ml lysozyme10% SDS4M NaOHautoclaved
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
Plasmid－isolation－from－yeast
Pick colonies into 0.5ml of SD-Leu (or other appropriate SD medium)Vortex for 1minLeave to grow O/N for 18-24h at 30°C, 230-250rpm (best in 5ml bijou)
质粒的大量制备
· Plasmid Mini and Maxi Prep Methods (Gimila Lab) · Maxi-preps and all media, solutions (NWFSC)Isolation of cosmid, plasmid and P1 DNA
质粒的大量制备
· Plasmid Mini and Maxi Prep Methods (Gimila Lab) · Maxi-preps and all media, solutions (NWFSC)Isolation of cosmid, plasmid and P1 DNA
Sauer:RNA－Purification－from－E.－coli
My Experience Purifying RNA from E. coliRegarding RNA extraction, there is a horrible tendency of people to use kits for RNA extraction with bacteria.
粘粒
Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re
粘粒
Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re
Plasmid－or－Cosmid－DNA－Miniprep
This protocol can be used to isolate sufficient amount DNA from 1.5ml o/n culture or 3ml 6hr culture to do several enzyme digestions.Spin 1.5ml o/n cu
Midiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
Screening－Bacterial－Colonies－Using－Xgal－and－IPTG:－αComplementation
实验概要 α-complementation occurs when two inactive fragments of E. coli β-galactosidase associate to form a functional enzyme. Many plasmid
plasmid－DNA的抽提•纯化试剂盒MagExtractor®Plasmid
产品索引 5870 中文名称： plasmid DNA的抽提•纯化试剂盒 英文名称： MagExtractor®-Plasmid- 产品编号： NPK -300 产品类别： 分子生物学 生产厂家： TOYOBO 产品价格：
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
DNA－Extraction－from－Tissue
实验概要DNA extraction from tissue.主要试剂Extraction buffer100 mM Tris-HCl (pH 8.0) 100 mM EDTA (pH 8.0) 100 mM Na-Phosphate (pH 8.0) 1.5 M NaCl1% CTAB
DNA－Extraction－from－Blood
实验概要The ChargeSwitch® gDNA Purification Kits allow rapid and efficient purification of genomic DNA from small volumes of human blood. After preparin
Quick－Yeast－DNA－Prep:－Isolation－of－Total－DNA－(genomic－and－plasmid)
Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg.Transfer culture to a small 13 x 100 glass tube. Spin down cells 2 min. in
Purifying－Large－E.－coli－Restriction－Fragments－from－PulsedField－Gels
DNA PreparationE. coli chromosomal DNA is prepared following the method of Heath et al. ( J. Bacteriol., 174, 1992). Cells are embedded in agarose, th
E.Z.N.A.™－Fastfilter－Plasmid－Mega－Protocol
实验概要The E.Z.N.A.™ family of products is an innovative system that radically simplifies extraction and purification of nucleic acids from a variety
DNA－Purification－from－Agarose－Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Isolation－of－genomic－DNA－from－bacteria
Note: This procedure does not work well with Gram + cocci.Transfer 1.5 mL overnight culture to a 1.5 mL microfuge tube, centrifuge for 30 sec, decant
Streptomyces:Protocols/Conjugation
Intergeneric Conjugation and OverlayDescription Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient str
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Maxipreparation
实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu
重组质粒(dna－recombinant－plasmid)的连接
质粒具有稳定可靠和操作简便的优点。如果要克隆较小的DNA 片段( ＜10kb) 且结构简单，质粒要比其它任何载体都要好。在质粒载体上进行克隆，从原理上说是很简单的，先用限制性内切酶切割质粒DNA 和目的DNA 片段，然后体外使两者相连接，再用所得到重组质粒转化细菌，即可完成。但在实际工作中，
One－step－miniprep－method－for－the－isolation－of－plasmid－DNA
plasmid miniprepAll ''miniprep'' methods reported so far for the isolation of plasmid DNA involve multiple pipetting, extraction, cent
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Minipreparation
实验概要Plasmid DNA is isolated from small-scale (1-2 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and Solutions: Alkaline lysis
重组DNA的分离、克隆与测序实验手册3
G. Bacterial cell maintenanceFour strains of E. coli are used in these studies: JM101 for M13 infection and isolation (4), XL1BMRF' (Stratagene) f
Cloning－PCR－products－using－TA－vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D. *Methods and reagents is a unique monthly column that highlights current discussions in