Protocol for Agrobacterium-mediated transformation in rice. Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2–3 months with a transformation frequency of 30–50%, both in easily cultured varieties and recalcitrant elite japonica rice. - [Read Agrobacterium-Mediated Transformation in Rice Protocol]
Electrotransformation of Agrobacterium with a plasmid that has been replicating in E. coli. Growth of Arabidopsis thaliana. Arabidopsis dunking. Seed Harvesting. Plant tissue culture. Very detailed protocol. Stockinger lab. PDF - [Read Arabidopsis transformation with Agrobacterium PDF]
Arabidopsis transformation and selection on kanamycin agar, Preparation of plants for transformation, Agrobacterium, Dipping, Post-dip care, Agar plates, Tips. Adaptation of the short protocol described bySteve Clough and Andrew Bent, University of Illinois at Urbana-Champaign. - [Read Arabidopsis transformation with pictures]
4 strains of E. coli are used in these studies: JM101 for M13 infection and isolation, XL1BMRF'for M13 or pUC-based DNA transformation, and ED8767 for cosmid DNA transformation. To maintain their respective F' episomes necessary for M13 viral infection, JM101 is streaked onto a M9 minimal media plate and XL1BMRF' is streaked onto an LB plate containing tetracycline. ED8767 is streaked onto an LB plate. These plates are incubated at 37degC overnight. For each strain, 3 ml. of appropriate liquid.. - [Read Bacterial Cell Maintenance Protocol]
Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding. - [Read C. elegans RNAi Protocol]
Cloning protocols/tips by Astrid. PCR and primer design, Extracting DNA from gel, Restriction of vector, Ligation, and Transformation. CalTech - [Read Cloning protocols/tips by Astrid PDF]
Polybrene and DMSO can be used to achieve stable transformation of several types of cells by plasmid DNA. The yield of transformants is up to 15-fold greater with Polybrene than with calcium phosphate-DNA coprecipitation. However, there is no difference between the two methods in the efficiency of transformation of cells by high-molecular-weight DNA. - [Read DNA Transfection Using Polybrene Protocol]
Protocol for Epstein-Barr Virus transformation of lymphoblasts. Protocol describes a method for the transformation of lymphoblasts by Epstein-Barr Virus (EBV). Cells may be isolated from whole blood or taken from cryopreserved, non-immortalized stocks. - [Read Epstein-Barr Virus Transformation of Lymphoblasts Protocol]
Expression of F-tryptophan labelled protein. Protocol can be adapted for Fluoro-phenylalanine or Fluro-tyrosine labelling. Transformation of competent E. coli cells for expression with plasmid. Dr. Chen, Dept. of Biochem. & Mol. Biology, University College, London. - [Read Expression of F-tryptophan labelled protein]
Protocol for fission yeast media. Includes: YES (yeast extract with supplements) for rich complete media; YSO, for modified rich media; EMM (Edinburgh minimal medium) for minimal selective media; stock solutions to make up minimal media; MB, a very stringent minimal for transformation; Sporulation media; ME (malt extract); SPAS; EMMGlut. - [Read Fission Yeast Media Protocol]
The plant transformation procedures described involve floral dip, vacuum infiltration, and spraying. They yield transformants at frequencies ranging up to several percent, with the most common frequency being 0.1%-1%. - [Read In Planta Transformation of Arabidopsis Protocols]
Protocol for large-scale yeast transformation. Includes: Yeast Cell Preparation; Large Scale Transformation; To plate on solid medium; To select in liquid. - [Read Large-Scale Yeast Transformation Protocol]
Ligation with GUS vector and Transformation. Chemical Transformation of E. coli. Ligation Procedure for LigaFastâ„¢ Rapid DNA Ligation System (Promega). B. Beason, Rice University. - [Read Ligation with GUS vector and Transformation]
The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.
This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.