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Hello everyone! I am currently using different DNA isolation kits to extract DNA from insects. Even though I am able to successfully extract DNA I would like to maximize the yield. Do you have any tips that might help me with that even if the kits are not specifically designed for insect samples?

Discussions What DNA isolation kit would work for insect samples?

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - BHK-21

Products Sigma-Aldrich CelLytic™ NuCLEAR™ Extraction Kit

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - BHK-21

Products Thermo Fisher Scientific RIPA Lysis and Extraction Buffer

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - Rat_Mesenteric fat

Products Sigma-Aldrich CelLytic™ MT Cell Lysis Reagent

Get tips on using AllPrep DNA/RNA Mini Kit to perform AAA for reviews

Products Qiagen AllPrep DNA/RNA Mini Kit

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - HUVEC

Products Sigma-Aldrich CelLytic™ MT Cell Lysis Reagent

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - HeLa

Products Sigma-Aldrich CelLytic™ MT Cell Lysis Reagent

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 Mt-PA

Products Thermo Fisher Scientific Expi293™ Expression System Kit

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Cellular assays ROS assay cell type human umbelical vein endothelial cells (HUVEC)

Protein expression refers to the techniques in which a protein of interest is synthesized, modified or regulated in cells. The blueprints for proteins are stored in DNA which is then transcribed to produce messenger RNA (mRNA). mRNA is then translated into protein. In prokaryotes, this process of mRNA translation occurs simultaneously with mRNA transcription. In eukaryotes, these two processes occur at separate times and in separate cellular regions (transcription in nucleus and translation in cytoplasm). Recombinant protein expression utilizes cellular machinery to generate proteins, instead of chemical synthesis of proteins as it is very complex. Proteins produced from such DNA templates are called recombinant proteins and DNA templates are simple to construct. Recombinant protein expression involves transfecting cells with a DNA vector that contains the template. The cultured cells can then transcribe and translate the desired protein. The cells can be lysed to extract the expressed protein for subsequent purification. Both prokaryotic and eukaryotic protein expression systems are widely used. The selection of the system depends on the type of protein, the requirements for functional activity and the desired yield. These expression systems include mammalian, insect, yeast, bacterial, algal and cell-free. Each of these has pros and cons. Mammalian expression systems can be used for transient or stable expression, with ultra high-yield protein expression. However, high yields are only possible in suspension cultures and more demanding culture conditions. Insect cultures are the same as mammalian, except that they can be used as both static and suspension cultures. These cultures also have demanding culture conditions and may also be time consuming. Yeast cultures can produce eukaryotic proteins and are scalable, with minimum culture requirements. Yeast cultures may require growth culture optimization. Bacterial cultures are simple, scalable and low cost, but these may require protein specific optimization and are not suitable for all mammalian proteins. Algal cultures are optimized for robust selection and expression, but these are less developed than other host platforms. Cell-free systems are open, free of any unnatural compounds, fast and simple. This system is however, not optimal for scaling up

Proteins Protein Expression Prokaryotic cells E. coli A. phagocytophilum MSP2

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