transformants Protocols

Category: Transfection Protocols

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]

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the expression of cloned genes in E. coli using IPTG-inducible promoters. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying IPTG-inducible promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using IPTG-inducible Promoters Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage {lambda} pL promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol]

Category: E Coli Protocols: E coli Protein Protocols

Protocol for expression of cloned genes in E. coli using the bacteriophage lambda pL promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage lambda pL promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol]

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the expression of cloned genes in E. coli using the bacteriophage T7 promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage T7 promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage T7 Promoter Protocol]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

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]

Category: Yeast Protocols: Yeast Genetics Protocols

When more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt by interaction mating. In this protocol one strain is transformed with library DNA and the transformants are collected and frozen in aliquots. - [Read Performing a Hunt by Interaction Mating Protocol]

Category: DNA Protocols: TOPO Cloning

Topo Cloning procedure for cloning of small amounts DNA fragments by PCR. Cloning the PCR fragment into the TOPO vector, transforming E. Coli cells, and using blue/white selection to determine transformants. Culture inoculation, Qiagen Plasmid Prep protocol for plasmid isolation of the plasmid containing the cloned copies. Preuss Lab, Univ. Chicago. - [Read Topo Cloning Procedure]

Category: Bacteria Transformation Protocols

Protocol describes a method for transforming Agrobacterium with plasmid DNA using electroporation in a manner similar to that commonly used for Escherichia coli. Although the transformation efficiency for Agrobacterium is lower than that for E. coli, it is possible to obtain adequate numbers of Agrobacterium transformants with this technique. - [Read Transformation of Agrobacterium Using Electroporation Protocol]

Category: Bacteria Transformation Protocols

Protocol describes a method for transforming Agrobacterium with plasmid DNA using a freeze-thaw technique. Although the transformation efficiency for Agrobacterium is lower than that for Escherichia coli, it is possible to obtain adequate numbers of transformants with this technique. - [Read Transformation of Agrobacterium Using the Freeze-Thaw Method Protocol]

Category: Tetrahymena Protozoa Protocols

Protocol describes a method for transformation of the Tetrahymena using electroporation. The vector is electroporated into cells after mating, where it is incorporated into the DNA of developing macronuclei. Because T. thermophila can be propagated indefinitely without conjugation, transformation of the macronucleus provides a way to obtain stable somatic transformants. DNA vectors transformed using this protocol include those containing drug-resistant versions of Tetrahymena genes. - [Read Transformation of Tetrahymena thermophila by Electroporation Protocol]

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol describes a method for transformation of the Tetrahymena using electroporation. The vector is electroporated into cells after mating, where it is incorporated into the DNA of developing macronuclei. Because T. thermophila can be propagated indefinitely without conjugation, transformation of the macronucleus provides a way to obtain stable somatic transformants. - [Read Transformation of Tetrahymena thermophila by Electroporation Protocol]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for vacuum-infiltration transformation of Arabidopsis thaliana. Includes: Plant Growth; Vacuum Infiltration; Selection of Putative Transformants; Kanamycin selection. - [Read Vacuum-Infiltration Transformation of Arabidopsis thaliana Protocol]