Cloning and Expression

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Simplicon™ RNA Transfection and Electroporation Protocols

Restriction enzyme collection has been optimized for digestion using five unique buffers.

Dephosphorylation Procedures for DNA and Proteins

Dephosphorylation of DNA using Calf Intestinal Alkaline Phosphatase, Shrimp Alkaline Phospha, Bovine Intestinal Alkaline Phosphatase

x-tracta Gel Extraction Tool

A quick, clean and easy to use alternative to using a scalpel or razor blade for the extraction of DNA and RNA bands from agarose gels following gel electrophoresis.

Blue-White Select™ Screening Reagent

Blue/white color selection is a routine technique employed by molecular biologists. This technique simplifies the differentiation between colonies/plaques that contain a cloning vector without an insert and those that contain a vector harboring an insert of interest.

pGEX Vectors

This page describes the characteristics of pGEX expression vectors used with Cytiva products.

Escort™ III Transfection Reagent Protocol

The product bulletin providin detailed use protocol for easy DNA transfection.

QuickLink™ DNA Ligation Kit

T4 DNA ligase is used for the joining of DNA molecules with compatible cohesive (sticky) termini, joining of blunt ended double stranded DNA molecules

OverExpress™ Chemically Competent Cells

Protocol for OverExpress™ Chemically Competent Cells. Product Numbers: CMC0017, CMC0018, CMC0019, CMC0020, CMC0023, CMC0024

Alkaline Phosphatase Protocol

Alkaline phosphatase (AP) is a non-specific phosphomonoester hydrolase that catalyzes the hydrolysis of a wide variety of organic monophosphates.

Universal Transfection Reagent Protocol

Our Universal Transfection Reagent is a unique formulation of a proprietary polymer blend used for transient and stable transfection of nucleic acids into various eukaryotic cell lines and hard-to-transfect primary cells. This is a fast and easy protocol is compatible

Calf Intestinal (CIP) Alkaline Phosphatase for Nucleic Acid Dephosphorylation

CIP is used to remove 5’-phosphate groups from DNA, RNA and both ribo and deoxy-ribonucleoside triphosphates. Detailed protocol on how to dephosphorylate DNA.

Competent Cell Protocols

Technical Article on competent cells. Transformation is a process by which some bacteria take up foreign genetic material (naked DNA) from the environment.

Selecting Correctly Expressing Recombinants

Blue White Screening; DNA Minipreps; Screening by Restriction Digestion; Screening by PCR; Confirm cut plasmid sizes by agarose gel electrophoresis; DNA Maxipreps; DNA Precipitation; RNase Treatment; Clean-up of DNA

BL21(DE3) Electrocompetent Cells

BL21(DE3) Electrocompetent Cells are provided in 25 μL aliquots, sufficient for one reaction. Transformation is carried out in a 0.1 cm gap cuvette. Optimal settings for electroporation are listed in the table below. Note that alternate settings result in transformation

Next Generation Cell Free Protein Expression Kit (Wheat Germ) (CFPS700) PROTOCOL

Protein synthesis using cell-free protein synthesis reagent kit consists of three stages: preparation of transcription template, transcription and translation.

Klenow Enzyme Protocol

Co-transfection of Plasmid DNA

Transient co-transfection of plasmids is a method that is commonly employed for cellular protein-protein interaction studies, transcription factor studies, and gene knockdown studies using shRNA encoding plasmids.

Calcium Phosphate Transfection Kit Protocol

Calcium phosphate transfection is a common method for the introduction of DNA into eukaryotic cells. This protocol can be optimized for use with a wide variety of cell types.

Rapid DNA Ligation Kit Protocol

Preparing Cells for Electroporation

DNA Ligation Protocol

The cloning process requires the ligation of linear DNA into a cloning vector. This ability to join fragments of DNA through recombinant technology is essential for many basic experiments in biotechnology.

OverExpress™ Electrocompetent Cells

OverExpress™ Electrocompetent Cells are provided in 25 μL aliquots, sufficient for one transformation reaction.

Alkaline Phosphatase (AP) Protocol

Alkaline phosphatase (AP) is a non-specific phosphomonoester hydrolase that catalyzes the hydrolysis of a wide variety of organic monophosphates.

Reverse Transcription Protocol

Method for reverse transcription of RNA into DNA. Uses a premixed reagent that contains reverse transcriptase, dNTPs, primers, RNase inhibitor and buffer. Fast generation of cDNA.

SIG10 Chemically Competent Cells

For best results, ligation reactions must be heat inactivated at 70º C for 15 minutes before transformation. Alternately, the reactions may be purified.

X-tremeGENE™ 9 DNA Transfection Reagent Protocol

Plate cells approx. 24 hours before transfection making sure cells are at optimal concentration (70 – 90 % confluency).

X-tremeGENE™ HP DNA Transfection Reagent Protocol

Cell preparation for transfection Plate cells approx. 24 hours before transfection making sure cells are at optimal concentration (70 – 90 % confluency).

Protocols for Transfecting Common Cell Lines with X-tremeGENE™ Transfection Reagents

Restriction Enzyme Cloning Manual Buffer Recipes

TE Buffer; Elution Buffer; 10x Ligation Buffer; 0.5 M PIPES Buffer; Inoue Transformation Buffer