siRNA / RNAi /miRNA transfection Human Cells Jurkat cells

Get tips on using ON-TARGETplus Mouse Ddit4 (74747) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RGC-5 Ddit4

Get tips on using ON-TARGETplus Mouse Eif2ak3 (13666) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - CT26 Perk/Eif2ak3

Get tips on using ON-TARGETplus Mouse Casp8 (12370) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - CT26 caspase-8

Get tips on using SignalSilence® NF-κB p65 siRNA I #6261 to perform siRNA / miRNA gene silencing Rat - H9c2 NF-κB RelA (p65)

Get tips on using Flot1 Rat siRNA Oligo Duplex (Locus ID 64665) to perform siRNA / miRNA gene silencing Rat - NRCM Flot1

Get tips on using Flot2 Rat siRNA Oligo Duplex (Locus ID 83764) to perform siRNA / miRNA gene silencing Rat - NRCM Flot2

Get tips on using JetPrime to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

Get tips on using JetPrime to perform DNA transfection Mammalian cells - Immortalized cell lines COS7

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.

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 the 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.