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General plasmid ligation protocol. Adapt for experimental conditions.

The polyA Spin™ Kit is a rapid and convenient alternative to traditional column chromatography for the isolation of full-length poly(A)+ (mRNA) from cytoplasmic or "total" RNA isolated from eukaryotic tissue and cells. Poly(A)+ RNA selection is made by affinity chromatography using spin columns. Prepared essentially by the method of Gilham (1), this solid support has found widespread ...

β-Agarase cleaves the agarose subunit, unsubstituted neoagarobiose [3,6-anhydro-a-L-galactopyranosyl-1-3-d-galactose] to neoagaro-oligosaccharides (1).
Source:
Isolated from a strain of E. coli that carries a plasmid which encodes the β-Agarase I gene.
Applications:
β-Agarase I digests agarose, releasing trapped DNA and producing carbohydrate molecules which...

Homemade protocol, no kit is used Please check the buffer section first. Obtained from www.protocolsexchange.com

Table for Method 1: Tube
Water (ml) Sample (ml) Glucose Standard (ml) Reagent Blank 1.00 -- -- Standard # 1 0.98 -- 0.02 Standard # 2 0.96 -- 0.04 Standard # 3 0.94 -- 0.06 Standard # 4 0.92 -- 0.08 Test -- 1.00 -- Table for Method 2: Tube Water (ml) Sample (ml) Glucose Standard (ml) Reagent Blank 1.00 -- --...

The antigen is extracted from the cell in an appropriate lysis buffer, and antibodies are added to the lysate to allow formation of the immune complex. A solid phase matrix containing Protein A or G is added, and the immune complexes are allowed to bind by adsorption of the antibody to Protein A or G. After the Protein A (orG)-antibody interaction occurs, the unbound proteins are removed ...

Antibodies used for cell surface labeling were purchased from BD PharMingen (San Diego, CA) and included fluorescein isothiocyanate (FITC)–labeled anti-Thy-1.2, anti-Gr-1, and anti-bromodeoxyuridine (BrdU), phycoerythrin-labeled anti-CD4, anti-CD3, and anti-NK1.1. Biotinylated anti-CD3ε, anti–T-cell receptor (TCR)αβ, anti-Thy-1.2, and anti-Mac-1 (CD11b) were detected by streptavidin-Cy5 (Jackso...

A simple protocol to follow, don’t forget the use DDW. Suits for most of plant tissues.

Homemade protocol, no kit is used Please check the buffer section first. Obtained from www.protocolsexchange.com

The CelLytic™B Plus Kit contains all of the reagents and chemicals necessary to lyse both Gram-negative and difficult to lyse Gram-positive bacteria. The kit also includes protease inhibitors to help prevent the proteolytic breakdown of proteins. The method provided can be used to extract soluble proteins, and can be used to remove cellular debris from inclusion bodies to yield nearly pur...

These instructions are designed for a 96 well plate format. If the assay is performed in a larger format, the volumes should be increased accordingly.Notes: It is recommended to add a negative and a positive control for each experiment:
•  A negative control will indicate the background level. A medium, which was incubated at 65 °C, with no secreted AP can serve as a negativ...

Having knowledge of the entire 3′ sequence of a cDNA is often important because the non-coding terminal region can contain signals that regulate the stability or subcellular localization of the mRNA. Also, some messages use alternative genomic sites for cleavage and polyadenylation that can alter the above properties, or change the encoded protein. Full-length cDNAs can be obtained from c...

ProcedureOne procedure is provided for processing fresh, frozen, RNAlater-preserved, and/or FFPE tissues. The WGA process is divided into Lysis and Fragmentation, OmniPlex Library Generation, and PCR Amplification steps. Ideally these steps should be carried out sequentially without pause, as storage between steps may allow the ends of the library DNA to degrade, thus affecting subsequent steps...

The protocol established by Kim et al. 2006.
Materials - Strain BL21 (DE3) E. coli cells harboring the Aβ42-GFP fusion vector.LB media supplemented with 50 μg/mL kanamycin. Concentrations of EM-1 and Aib-1 Concentration EM-1...