- Ideal even for difficult-to-lyse Gram positive bacteria
- Yields the highest quality bacterial total RNA
- No enzymatic pretreatment required
- DNA-free™ reagents included for RT-PCR ready RNA
The steep rise in the number of antibiotic resistant bacterial species has led to an intensive effort to develop designer antibiotics and other therapeutic agents to contain this growing threat. This in turn has created a need to dissect bacterial genomes and understand bacterial gene expression pathways in greater detail. Array analysis, the best tool yet available to monitor global changes in gene expression, requires the isolation of high quality RNA. Traditionally, the isolation of total RNA from bacteria has required the use of hot phenol or other such harsh techniques. If the microbe being studied is a Gram positive organism, pretreatments with cell wall digesting enzymes might also be required prior to isolation of RNA. However, since these enzymatic pretreatments are thought to alter gene expression patterns on a global scale, array data can be compromised.
The RiboPure-Bacteria RNA Isolation Kit uses no enzymatic pretreatments and thus does not alter expression profiles. RiboPure-Bacteria combines an efficient glass bead mediated organic disruption step followed by glass filter based RNA purification for high yields of exceptionally pure bacterial RNA. DNase treatment can be performed with the provided DNA-free™ reagents for DNA sensitive applications such as RT-PCR. Additionally, Ambion's RNAlater® RNA Stabilization Solution can be used to store intact bacteria for days or weeks prior to RNA isolation without compromising the integrity of the RNA. As seen in the lower figure at right, RNA isolated from Bacillus subtilis using the RiboPure-Bacteria Kit was stored for a week at 4ºC in RNAlater is of very high quality compared to RNA isolated from bacteria stored in TE in the same manner. The kit has been used to isolate high quality RNA from Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Campylobacter fetus, and Rhodobacter sphaeroides.
The RiboPure-Bacteria RNA Isolation Kit contains sufficient reagents for 50 RNA isolations, each from 108 to 109 cells. The Kit requires the use of the Ambion Vortex Adapter (which turns a standard laboratory vortex into a bead beater). This adapter fits the Vortex Genie® 2 and holds 12 tubes.
For applications where the use of mRNA is desirable, this kit can be used in conjunction with the MICROBExpress™ Bacterial mRNA Enrichment Kit.
Biocompare® Product Review
Trademarks, Patents, Licensing
|
