Protein Ladder

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 pLEXSY-hyg-2-hAm to perform Protein Expression Eukaryotic cells - Iranian lizard Leishmania cells recombinant human amelogenin

Get tips on using pPZP-RCS2-ocs-nptII to perform Protein Expression Prokaryotic cells - A. tumefaciens α-amylase, amylopullulanase, and glucoamylase

Get tips on using MicroVue YKL-40 EIA to perform ELISA Human - Chitinase-3-Like Protein-1 (CHI3L1) or YKL-40

Get tips on using pYT379-CDK8-CycC-10xHis complex to perform Protein Expression Eukaryotic cells - S. frugiperda CDK8-CycC-10xHis complex

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

Get tips on using pET30 Ek/LIC-Colα3(IV) NC1 to perform Protein Expression Prokaryotic cells - E. coli Colα3(IV) NC1

Get tips on using pPICZα A, B, & C Pichia Vectors to perform Protein expression and purification Yeast - Pichia pastoris EDIII-D1

Get tips on using pcDNA™3.1 (+) Mammalian Expression Vector to perform Protein expression and purification Mammalian cells - CHO-K1 sRAGE