siRNA / RNAi /miRNA transfection Mouse Primary Splenocytes

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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 Anti-Estrogen Receptor (ER) (SP1), Rabbit Monoclonal Primary Antibody to perform Immunohistochemistry Estrogen receptor (ER) - Rabbit Human -NA-

Get tips on using ON-TARGETplus Rat Smad2 (29357) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad2

Get tips on using ON-TARGETplus Rat Smad3 (25631) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad3

Get tips on using ON-TARGETplus Rat Hspa5 (25617) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - AR42J Grp78/Hspa5

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Get tips on using ON-TARGETplus Rat Atg7 (312647) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Atg7

Get tips on using ON-TARGETplus Rat Becn1 (114558) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Becn1