siRNA / miRNA gene silencing Human PANC-1

Get tips on using claudin-1 Antibody (XX7): sc-81796 to perform Western blotting Cclaudin-1

Get tips on using Mouse Dkk-1 Quantikine ELISA Kit to perform ELISA Mouse - Dkk-1

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - immortalized A-172

Get tips on using PAXgene Tissue miRNA Kit to perform RNA isolation / purification Tissue - rat heart muscle tissue

Get tips on using Beclin-1 Antibody, ProSci to perform Autophagy assay cell type - IEC-6

Get tips on using LAMP-1 Polyclonal Antibody to perform Autophagy assay cell type - RAW 264.7

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 pSUPER.retro.puro to perform shRNA gene silencing Rat - MM1 SSH1

Get tips on using pSUPER.retro.puro to perform shRNA gene silencing Rat - MM1 SSH2

Get tips on using pSUPER.retro.puro to perform shRNA gene silencing Rat - MM1 LIMK1