PCR/Cloning Reagents

Competent cells

Xtreme Efficiency Competent Cells are among the most commonly used E. coli strains for plasmid transformation. They are prepared with unique salt compositions and procedures that result in significantly higher transformation efficiencies than those prepared by traditional lab protocols using CaCl2 buffers. The cells are also quality controlled by direct comparison to other leading brands in the area of transformation of ligated plasmids. Chemically competent DH5α cells are suitable for a short transformation protocol in as little as 3 minutes before plating. Also, the cells are suitable for the propagation of plasmids of all sizes and preparations, and are recommended for difficult constructs that often yield low colony counts. They compare favorably to anything you will find in the market, and are about half the price of those from most well known sources.

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Cloning enzymes

Our cloning enzymes, made available through the PureAct series, are prepared by non-traditional purification procedures. As a result, the enzymes such as T4 polynucleotide kinase, ligase, and polymerase are highly active and pure. They catalyze the intended reactions without any detectable side reactions, such as degradation, even after long reaction times.

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PCR Cloning

The Allele TA vector is a high quality cloning vector used to facilitate the cloning of PCR products. This vector has been linearized and is flanked by many recognition sites for restriction enzymes. The vector possesses single 3’ thymidine overhangs at the insertion site, which improves the ligation efficiency of PCR products into the vector due to the prevention of recircularization and gives the 3’ deoxyadenosine on the PCR product a matching overhang.

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Spreading Beads

The most efficient way of plating competent cells is to use autoclaved, single-use glass beads. Simply pour about 10 beads per 10-cm agar dish. Move the dish sideways a few times, and the transformed cells will be evenly distributed throughout the plate surface. Multiple plates can be plated at once. The spreading beads are specifically designed and tested for this purpose.

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One-Step Genotyping

One-Step Genotyping kits can provide genomic DNA for direct PCR detection from mouse tail or blood samples using a single reagent and a one temperature reaction in as little as 15 minutes making the kit a one of a kind in this product class. Additional kits for other sample types will become available soon. To facilitate or speed up lysis of samples, try the Allelequip incubator-shaker line of products.

Allele-In-One Mouse Tail/Human Blood Genotyping System

This system is based on our specifically formulated Allele-In-One Mouse Tail Direct Lysis Buffer, Allele-In-One PCR mastermix, and high performance Genomic DNA Purification System making them your ideal tools for quick, convenient, low cost, yet high-quality genotyping research. The direct lysis buffer contains a combination of enzymes, detergents, and other chemical reagents that will lyse the mouse tail tissues or human blood cells, releasing genomic DNA in a one-step reaction (incubate at 55oC for 2 hrs). After a brief centrifuge, the supernatant can be used directly as a template in a genotyping PCR reaction. No phenol extraction, precipitation, or further purification is necessary.

The Allele-In-One PCR mastermix is suggested to be used in combination with the lysis buffer, which is specifically formulated to accommodate DNA template generated from the lysis buffer containing proteases. However, you can still optimize your current PCR system for use with the lysis buffer. Many of our customers have had success doing so.

For certain experiments that require purified genomic DNA, e.g. SNP assays, the Allele Genomic DNA Purification System is recommended after lysing the mouse tail samples with the direct lysis buffer. This buffer can also be used to prepare genomic DNA for direct PCR from other tissues such as mouse ear, rat tissues, etc. Slight adjustments and optimization may be needed for each type of tissue.

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PCR enzymes & Reagents

If for any reason, you prefer to assemble your own PCR reaction rather than use Mastermix, we also offer certified Taq DNA polymerase, PCR buffers, dNTP solutions, dATP, dGTP, dCTP, and dTTP.

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