siRNA / RNAi /miRNA transfection Human Cells HESC

Get tips on using ON-TARGETplus Mouse Mapk1 (26413) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 MAPK1 (ERK2)

Get tips on using ON-TARGETplus Mouse Mapk14 (26416) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 p38/Mapk14

Get tips on using ON-TARGETplus Mouse Nr1h3 (22259) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - M210B4 LXR‐α/Nr1h3

Get tips on using ON-TARGETplus Mouse Nr1h2 (22260) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - M210B4 LXR‐β/Nr1h2

Get tips on using ON-TARGETplus Mouse Zbp1 (58203) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 3T3-SA Zbp1/Dai

Get tips on using Silencer® Select_FPr1/Silencer® Select_FPr2 siRNA to perform siRNA / miRNA gene silencing Mouse - Neuro 2a Fpr1/Fpr2

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

Get tips on using Slc1a2 to perform siRNA / miRNA gene silencing Rat - Glial cells GLT-1