REDExtract-N-Amp™ Tissue PCR Kit From Sigma-Aldrich

REDExtract-N-Amp™ Tissue PCR Kit From Sigma-Aldrich
REDExtract-N-Amp™ Tissue PCR Kit is a good kit in my opinion. I am a graduate student. My research involves characterizing the role of two proteins using an autoimmune mouse model. I have the gene for each protein knocked-out in individual mice. I am currently in the midst of understanding each protein and its respective contributions to a particular autoimmune disease. I am also interested in determining if there is a synergistic effect when both proteins are lacking. Thus, I am currently breeding the two knockouts to get double knockouts. Hence I use the REDExtract-N-Amp™ Tissue PCR Kit quite a bit. While I have not used any other kit or method for genotyping my mice I feel like this kit is a dependable method of genotyping. The REDExtract-N-Amp™ Tissue PCR kit was actually recommended to me by an animal technician from another lab.

The REDExtract-N-Amp™ Tissue PCR Kit is, if anything, complete. It contains a tissue preparation solution and extraction solution. In my protocol, I mix and aliquot these two solutions into tubes, then cut the tails into the tubes. By the time I return from the animal facility, the 10 minute incubation at room temperature is done. Next, I pop the tubes into a 95°C dry bath for 3 minutes, add the neutralization buffer, then I am PCR-ready. Speaking of PCR-ready, the loading dye is included in the master mix provided in the kit. The master mix can be bought independent of the kit, which is convenient when you are looking for two different genes from the same sample. Personally, it really bothers me when you have to buy the entire kit to replace one element! Even if you do have to replace the kit, the prices are reasonable.

This specific kit allows for amplification of genomic DNA extracted from mouse-tails (0.5 to 1.0 cm), hair (1 root), tissue (2 mg to 10 mg), and buccal swabs (10 ul). Sigma has other similar kits for extraction from whole blood, plant leaves, and plant seeds.

The bad news, while using the kit, my master mix did go bad. Sigma guarantees the products for only one year. If you do not genotype frequently, but you want to buy in bulk to reduce cost you may end up paying more if the master mix goes. Also, if you run out of the master mix provided by the kit, I should tell you that this kit is optimized for use with its own master mix. Use of another PCR master mix, or making up of your own master mix will take some optimization. Even with optimization, the PCR products may not be clear, or completely absent. However, if everything is there and the protocol is performed as suggested, it works beautifully every time.

In summary, this is definitely a great product. I would not expect anything less from Sigma. Most importantly, this kit is quick, complete, and extremely dependable. The master mix can go bad before you use the entire kit. I would definitely recommend the REDExtract-N-Amp™ Tissue PCR Kit and I appreciate that it was recommended to me. Definitely saves time.

Graduate Student
Department of Microbiology and Immunology
Temple University
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REDExtract-N-Amp™ Tissue PCR Kit From Sigma-Aldrich
The Good

From tissue to PCR-ready in 15 minutes!

The Bad

Master mix may expire before you use the entire kit.

The Bottom Line

If you are genotyping on a small or large scale, use this kit! Save the time.

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