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

Methods-for-the-Measurement-of-a-Bacterial-Enzyme-Activity-in-Cell-Lysates

AbstractThe kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of Helicobacte

TritonPrep-Method-for-bacterial-DNA-Purification

Triton-Prep Method for bacterial DNA PurificationGrow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube).Resus

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

PEGMediated-Protoplast-Transformation-with-Naked-DNA

Direct introduction of DNA into plant protoplasts facilitates a rapid analysis of transient gene expression, as well as the generation of stably t

超级感受态细胞的制备

The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" CellsJoseph SambrookPeter Maccallum Cancer Institute and T

Preparation-of-Bacterial-Proteins-for-Analysis-by-2DPAGE

The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ

Construction-and-Manipulation-of-LargeInsert-Bacterial-Clone-Libraries

Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim

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

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

Methods-for-the-Measurement-of-a-Bacterial-Enzyme-Activity-in-Cell-Lysates4

The use of a micro-titre based colorimetric assay provided a third method for the study of ACTase activity, and the most useful for large scale measur

Methods-for-the-Measurement-of-a-Bacterial-Enzyme-Activity-in-Cell-Lysates2

Measurement of ACTase activityNuclear magnetic resonance spect roscopy (NMR)The unique potential of NMR spectroscopy for monitoring simultaneously the

Methods-for-the-Measurement-of-a-Bacterial-Enzyme-Activity-in-Cell-Lysates3

Different enzyme assays for ACTase study in H. pyloriACTase properties were studied in situ in cell-free extracts to obtain information on enzyme func

细菌培养基

Preparation of LB Plate (Dr. Chastain)prepare LB plate with or without antibioticsBacterial Culture Media Recipes (WUGSC) M9 Plate Supplement (Gottsch

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

Bacterial-Expression-of-IRS1-containing-GSTfusion-Proteins

1.  Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2.  Grow larger culture using the overnight culture as a seeding culture. 

Genetic-Transformation-of-Cotton-with-a-HarpinEncoding-Gene-hpaXoo-Conf...

Genetic Transformation of Cotton with a Harpin-Encoding Gene hpaXoo Confers an Enhanced Defense Response Against Verticillium dahliae KlebThe soil

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

DNA克隆

DNA克隆(主要内容如下)·         General Procedure·         PCR Cloning·         Subcloning·         ET Cloning·         Vector Preparation·         Ligation Re

DNA转化实验指导3

6.     Simultaneous digestion of the pUC vector with both enzymes in the presence of 3 units of Shrimp Alkaline Phosphatase (Amersham BioSciences) in

TOP10-chemically-competent-cells

OverviewThis protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the B

Basic-procedures-for-bacteria-culture2

E. Elution of DNA fragments from agaroseDNA fragments are eluted from low-melting temperature agarose gels using an unpublished procedure first develo

重组DNA的分离、克隆与测序实验手册6

Electroporation ProtocolPreparation of Electro-competent Cells:1. Grow XL1-Blue cells on a tetracycline plate (20 ug tet/ml of LB agar)2. Inoculate 3

DNA转染

DNA转染·         Transfection of Mammalian Cells Using Lipofectamine (LTI)·         Guide to Eukaryotic Transfections with Cationic Lipid Reagents (PDF)

E.Z.N.A.®-Plasmid-Maxi-Kit-vacuum-Protocol

实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c

E.Z.N.A.®-Plasmid-Maxi-Kit-Spin-Protocol

实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c

酵母转化

·         Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation·         Yeast Transformation (Breeden Lab)LiAc method·         Large-Scale Y

Fastfilter-Plasmid-Midi-Kit-Vacuum/Spin-Protocol

实验概要The E.Z.N.A.TM Fastfilter Plasmid Midi Kit combines the power of HiBind® technology with the time-tested consistency of alkaline-SDS lysis of

Fastfilter-Plasmid-Midi-Kit-Spin-Protocol

实验概要The E.Z.N.A.TM Fastfilter Plasmid Midi Kit combines the power of HiBind® technology with the time-tested consistency of alkaline-SDS lysis of

重组DNA的分离、克隆与测序实验手册5

C. Random fragment end-repair, size selection, and phosphorylationSince both sonicated and nebulized DNA fragments usually contain single-stranded end

离心的基础知识

Centrifugation is a process used to separate or concentrate materials suspended in a liquid medium. The theoretical basis of this technique is the eff