RNAprotect Bacteria Reagent
For stabilization of RNA in bacterial cultures
RNAprotect Bacteria Reagent provides immediate stabilization of RNA prior to RNA isolation procedures, thereby ensuring reliable gene expression and gene-profiling data. Adding the reagent directly to bacterial cultures achieves stabilization of both Gram-positive and Gram-negative bacteria.
Use of RNAprotect Bacteria Reagent overcomes the problems of RNA degradation and changes in expression profiles by providing immediate stabilization prior to RNA isolation (see figures "RNAprotect Bacteria Reagent prevents mRNA degradation" and "GeneChip analysis"). RNAprotect Bacteria Reagent is suitable for use with a wide range of bacterial species, both Gram positive (e.g., Staphylococcus aureus and Mycobacterium avium) and Gram negative (e.g., Escherichia coli and Salmonella typhimurium). Bacteria can be grown in either minimal or complex medium prior to isolation of RNA.
Traditional methods for cell harvesting and RNA isolation can lead to vast changes in bacterial expression profiles. Enzymatic degradation of RNA leads to reduction or loss of many transcripts because bacterial mRNA molecules have very short half lives of only a few minutes. In addition, genes can be induced during handling and processing of samples, leading to higher expression levels of certain transcripts. Use of RNAprotect Bacteria Reagent overcomes these problems by providing immediate stabilization of RNA prior to RNA isolation procedures.
Two volumes of reagent are added directly to 1 volume of bacterial culture prior to RNA isolation, providing immediate stabilization of RNA (see figure "RNAprotect Bacteria Reagent procedure"). The stabilization allows time for efficient bacterial lysis using a choice of protocols: enzymatic lysis, mechanical disruption, or a combination of both methods. QIAGEN recommends the TissueLyser for efficient mechanical disruption.
Bacteria stabilized in RNAprotect Bacteria Reagent can be used for total RNA isolation using RNeasy Kits.
fragment fix placeholder