RNAlater (50ml)，76104，Qiagen，凯杰

RNAprotect Reagents and Tubes

For immediate stabilization of RNA in tissues, cultured cells and bacterial cultures

Features

Immediate stabilization of RNA and RNA protection

Convenient and safe handling at room temperature

Easy collection and storage of tissues in reclosable tubes

Stabilization of Gram-positive and Gram-negative bacteria

No need to remove medium from sorted or cultured cells

RNAprotect Reagents and Tubes provide immediate RNase inactivation and stabilization of RNA in tissues, cultures of Gram-positive and Gram-negative bacteria and sorted or cultured cells to preserve the gene expression profile and enable reproducible purification of high-quality RNA. Samples are stabilized at room temperature prior to RNA isolation procedures.

RNAprotect Tissue Reagent ensures immediate RNase inactivation and stabilizes and protects the RNA expression pattern of samples during harvest and storage (see figure " RNAprotect Reagent prevents degradation of mRNA in tissues"). Immediate protection with RNAprotect technology ensures that downstream analyses truly reflect the expression profile of the intact tissues. Samples can be archived without risk of RNA degradation, even after multiple freeze–thaw cycles (see figures “ Stable RNA in tissues: different temperatures and freeze–thaw cycles” and “ RNAprotect Tissue Tubes prevent RNA changes in tissues”).

RNAprotect Cell Reagent prevents induction of gene expression in treated cells and preserves and stabilizes transcript levels during storage of cell samples (see figures " Effective inhibition of PMA induction of c-fos expression" and " More reliable quantification of CD83 transcript").

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) and prevents RNA changes (see figures “ RNAprotect Bacteria Reagent prevents mRNA degradation” and “ GeneChip analysis”). Bacteria can be grown in either minimal or complex medium prior to isolation of RNA.

GeneChip analysis.

RNeasy purified RNA from RNAprotect stabilized samples stored under a wide variety of conditions is ideal for downstream processes such as northern blotting and RT-PCR.

Stable RNA in tissues: different temperatures and freeze–thaw cycles.

Total RNA was isolated from tissue samples stored in RNAprotect Tissue Reagent. [A] Rat lung tissue was stored under the indicated conditions. RNA was analyzed on a formaldehyde agarose gel and northern blot (GAPDH-probed). [B] Stabilized mouse liver tissue was subjected to freeze–thaw cycles (-20°C/25°C). C: control tissues frozen in liquid nitrogen and stored at -80°C.

Effective inhibition of PMA induction.

Incubation of PMA-stimulated Jurkat cells in RNAprotect Cell Reagent prevents PMA induction of c-fos expression as well as degradation of c-fos transcript, demonstrating stabilization of RNA by RNAprotect Cell reagent (each bar represents the mean of duplicate cell samples).

RNAprotect Bacteria Reagent prevents mRNA degradation.

In order to monitor mRNA degradation only, transcription was stopped by adding the RNA polymerase inhibitor rifampicin to a growing culture of E. coli. The culture was split into halves, and RNAprotect Bacteria Reagent was added to one half. Samples were left at room temperature for 0, 4, 8, and 16 minutes before centrifugation and RNA isolation. The resulting RNA was analyzed by agarose gel electrophoresis (top panel). The expression of two marker genes with different half-lives was examined by northern blot analysis. Middle panel: ompA (half life of 15 minutes); bottom panel: α-lactamase (half life of 2–5 minutes).

RNA stabilized in RNAprotect Reagents and Tubes can be used for:

• Total RNA purification

• miRNA purification

• miRNA purification