Which brand of reagent kit is more stable for extracting genomic DNA?

Fluctuations in concentration and purity may not necessarily be a brand issue, first check your own operations. It is recommended to fix the sample size, lysis time, and centrifugation speed, and perform negative control and one known sample each time. A260/280 is measured to be around 1.8, and A260/230 is ideal when it is greater than 1.8-2.0. If A260/230 is too low, it indicates salt or guanidine salt residue, and washing frequency can be increased or air drying can be extended. The tail of a mouse contains a lot of connective tissue, so after digestion, centrifuge the supernatant to avoid sucking up the sediment. The phenol chloroform method is generally not required unless downstream is extremely sensitive to purity.

The stability of QIAGEN DNeasy Blood&Tissue imported is indeed good, and it is suitable for mouse tail and cell precipitation. However, the price is usually high, and a budget of 500-800 may only be able to buy small specifications or require promotion. Invitrogen PureLink and Promega Wizard are also stable, and the column membrane method does not require phenol chloroform. The main differences between imported and domestic products are the uniformity of the silicone membrane, removal of protein and salt residues, and extraction time of approximately 2-3 hours. If the budget is tight, we can import small specifications for comparison and use domestic large packaging for routine extraction.

Domestic Ritengen, Novizan, omnipotent gold, Kangwei century, and Biyuntian have all been used, and the stability of mouse tail extraction is more tested and cleaved than cell precipitation. After the tail is cut, it is recommended to add fresh protease K, shake at 56° C overnight or at least 3-4 hours, and incomplete digestion is the most common cause of sudden high and low concentrations. Pay attention to the amount of cell precipitation, 1-5 × 10 ^ 6 is enough. Domestic kits generally do not use phenol chloroform, and the column load and elution volume should be clearly seen. The yield can be improved by preheating to 60°C with TE or water during elution.

Budget 500-800 Buy 50-200 large packages made in China is more cost-effective, and imported formal wear may be tight. When asking for a quote, ask if it contains protease K and RNA enzymes, many of which are low in price but need to be prepared separately. The arrival period is usually 1-3 days for domestic production and 1-2 weeks for imports, which has little impact on routine experiments. It is recommended to apply for a trial package or small specification first, do parallel extraction on the tail and cell samples of the same batch of mice, compare the concentration, purity, and downstream PCR or enzyme digestion effects, and then decide on bulk procurement.