Ningbo Scientz Biotechnology Co.,Ltd. is one of the leading scientz ultrasonic homogenizer with soundproo box suppliers in China. Welcome to buy our quality and cheap products for sale in bulk. We also welcome wholesale orders and customized orders. Now, please be free to check the price list with our factory.
Welcome to buy our quality and cheap scientz ultrasonic homogenizer with soundproo box for sale in bulk. As one of the leading Ultrasonic Homogenizer suppliers in China, we also welcome wholesale orders and customized orders. Now, please be free to check the price list with our factory.
What are the conditions for sonication of Streptomyces (actinomycetes)?
Analytical: Actinomycetes belong to the (G+C) molar percentage (mol%) of the prokaryotic system phylogenetic tree
a branch of the genus Eubacteria, which has molecular and biological characteristics unique to prokaryotes,
However, in its different groups, the chemical composition of the cell wall varies greatly. In the E. coli ultrasound, 400W is used, and the method of breaking 5s for 5s is good, but it is used on Streptomyces, and it has no effect due to the difference in cell wall composition. The pre-spinning treatment of Streptomyces (actinomycetes) is generally configured to a certain concentration of bacterial suspension.
The specific conditions for the crushing using the ultrasonic cell disruptor are: collecting the bacteria in the 24 h culture solution at 5 000 r/min for 5 min to collect the cells; washing with the pH 7.5 Na2HPO-NaH2PO buffer for 3 times, and then using The buffer is formulated into a 1:3 bacterial suspension. Placed in a 40 mL large plastic test tube; place the large plastic test tube in an ice bath and crush it with an ultrasonic cell disrupter (power 200W, 1/2″ probe, crush for 30s, intermittent 30s); crushing solution at 12 000 r/min Centrifuge for 30 min at high speed, collect cell debris and supernatant. Wash cells can also be washed once with pre-cooled saline or Tris-HCl at pH 8. In addition, the ultrasonic dose varies with the amount of sample and bacteria. The power can be 400-600w, crushed for 5s, stopped for 5s, ice bath, and the final concentration of 1 mM PMSF is added. In order to determine the appropriate ultrasonic intensity and frequency, it is necessary to check whether the bacteria are completely broken at any time.
What I have to do is the release rate of LDH (lactate dehydrogenase) ie supernatant/cell + supernatant, but in the cell
Need to break the cells when LDH, what should you pay attention to when breaking?
Analysis: 1. Ultrasound should be carried out in an ice bath;
2. The ultrasonic power has a great relationship with the inner diameter of the container: the inner diameter is small, the required power is also small, and it is not easy to cause air bubbles;
Conversely, it may cause protein denaturation;
3. Adding some protease inhibitors before ultrasound is beneficial to protect the activity of the protein of interest;
4. Ultrasonic time 5′, clearance 15-20′, total time 20min or so.
