Navigation Links
Bacterial RNA Isolation From Infected Eukaryotic Hosts


Remove >90% of mammalian RNA from complex mixtures of host-bacterial samples

Simple procedure takes less than 2 hours

Works with any bacterial species

Seamless integration with MICROBExpress Bacterial mRNA Purification Kit

Enables microarray expression analysis with bacterial RNA isolated from host cells


Ambion's new MICROBEnrich (patent pending) is the only kit available for bacterial RNA enrichment from mixed host-bacterial RNA populations. A robust yet simple procedure, MICROBEnrich depletes mammalian (host cell) RNA selectively, leaving behind highly enriched bacterial RNA.

Gene expression analysis following host-bacterial interactions is revolutionizing our understanding of the host response to bacterial infection. Numerous studies have utilized microarray analysis to follow host cell transcriptome alterations in response to interactions with bacteria. However, because no method has existed to isolate bacterial RNA away from host cell RNA, the corresponding analyses of bacterial transcriptomes has been impossible. Ambion now introduces a new technology, MICROBEnrich, that enables whole genome microarray expression analyses of bacterial RNA after host-bacterial interactions.


Start With Any Total RNA Sample

Although the mass of eukaryotic RNA isolated from host-bacterial cell mixtures is often far greater than the mass of bacterial RNA isolated, this complex mixture of total RNAs is the starting material for the MICROBEnrich procedure. This host-bacterial cell total RNA mixture can be obtained by a variety of RNA isolation methods, ideally by Ambion's new RiboPure-Bacteria Kit. Other methods including glass fiber filter based methods (such as Ambion's RNAqueous Kits) and phenol based methods (such as Ambion's RNAwiz and the ToTALLY RNA Kit) are also appropriate. For microarray analysis and other sensitive applications, Ambion also recommends a DNase treatment (such as with DNAfree Treatment and Removal Reagents) to remove contaminating genomic DNA from the total RNA prior to the MICROBEnrich procedure.


Remove >90% of Mammalian RNA from Complex Total RNA Mixtures

MICROBEnrich employs hybridization capture technology (patent pending) to remove human, mouse, and rat RNA (both mRNA and rRNA) from complex host-bacterial RNA populations, leaving behind enriched microbial total RNA. In the first step of the MICROBEnrich procedure, host-bacterial total RNA is incubated with an optimized mixture of capture oligonucleotides that bind to the mammalian 18S and 28S rRNAs and polyadenylated RNAs. Next, the rRNA/oligo nucleotide hybrids and all polyadenylated mRNAs are removed from the mixture with oligonucleotide-derivatized magnetic beads. After magnetic separation, the bacterial RNA remaining in the supernatant and can be precipitated with ethanol. Figure 1 shows an Agilent 2100 bioanalyzer electropherogram of a complex host-bacterial RNA population before and after bacterial RNA purification with MICROBEnrich. The RNA profile indicates nearly complete removal (>95%) of the host 18S and 28S rRNAs.

Figure 1. RNA Before and After Enrichment with MICROBEnrich Kit. Rat liver total RNA (25 g) and E. coli total RNA (2 g) were mixed together and subjected to the MICROBEnrich procedure. Another mixed sample of total RNA was subjected to a mock MICROBEnrich procedure in which the capture oligonucleotide mix was left out of the reaction. RNA was precipitated and an equal volume of each sample was separated on an RNA Nano LabChip with the Agilent 2100 bioanalyzer.


Removal of Bacterial rRNA
If desired, upon completion of the MICROBEnrich procedure bacterial mRNA can be purified from the total RNA through the seamless integration of Ambion's MICROBExpress technology. This technology enables >95% removal of prokaryotic 16S and 23S rRNAs (Figure 2). See the article "Purify mRNA From Bacteria!" for information on MICROBExpress.

Figure 2. RNA Before and After Enrichment with MICROBEnrich. Rat liver total RNA (25 g) and E. coli total RNA (2 g) were mixed together and subjected to the MICROBEnrich procedure with or without a further enrichment by MICROBExpress. The total RNA lane was subjected to a mock MICROBEnrich procedure in which the capture oligo mix was left out of the reaction. RNA was precipitated and an equal fraction of each sample was analyzed by denaturing agarose gel electrophoresis.


MICROBEnrich Will Work With Any Bacterial Species
MICROBEnrich was optimized using E. coli RNA in a background of human, mouse and rat RNA. However, the kit should perform well with RNA from any bacterial species. Although many mammalian host species may be compatible with MICROBEnrich, only human, mouse and rat rRNA sequences were used for the design and optimization of the capture oligonucleotides.


Increased Sensitivity in Downstream Analyses

Bacterial mRNAs purified with the combined MICROBEnrich and MICROBExpress Kits serve as better templates than total RNA for downstream applications and therefore deliver dramatic increases in sensitivity when used in applications such as array analysis. Figure 3 shows Sigma-Genosys Panorama E. coli arrays that were hybridized with labeled cDNA derived from either a mixed population of total RNA (E. coli/HeLa cell RNA mixture) or mRNA purified from the mixture using the MICROBEnrich and MICROBExpress Kits. The number of transcripts detected with labeled RNA target increases substantially after bacterial mRNA enrichment using the MICROBEnrich and MICROBExpress Kits.

Figure 3. Enrichment for Bacterial mRNA Results in More Sensitive Array Analysis. Replicate portions of Sigma/Genosys E. coli Panorama arrays were hybridized under identical conditions with 33P-labeled cDNA synthesized from (A) HeLa cell RNA (10 g) mixed with E. coli RNA (800 ng) or (B) enriched E. coli mRNA that was purified from an identical mixture using MICROBEnrich and MICROBExpress Kits.


Reagents to Process 500 g of Host Cell RNA
The MICROBEnrich Kit comes with reagents capable of depleting 500 g of host cell RNA (20 rxns x 25 g RNA) from a complex host-bacterial RNA mixture. The Kit is scalable, and a single reaction can process between 10 and 100 g of total RNA. The Kit includes a control RNA sample (E. coli and rat liver total RNA) and a detailed Instruction Manual with tips and troubleshooting advice. A single place magnetic stand and Ambion's MICROBExpress Kit are available separately.


back to top


Ordering Information
For prices and availability, please contact our Customer Service Department.
Cat# Product Name Size 1901 MICROBEnrich Kit 20 purifications 1905 MICROBExpress Bacterial mRNA Enrichment Kit 20 rxns 5750 MICROBExpress Module: Pseudomonas 20 purifications 5751 MICROBExpress Module: Campylobacter/Helicobacter 20 purifications 5752 MICROBExpress Module: Rhodobacter 20 purifications 10026 Single Place Magnetic Stand 1 ea
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. High-Throughput Cultivation of Bacterial Clones
2. Mammalian and Bacterial Expression in One Vector
3. Bacterial mRNA Isolation, Fast and Easy
4. Bacterial RNA Isolation From Infected Eukaryotic Hosts
5. Bacterial Whole Genome Array Analysis
6. Perform Microarray Analysis with Limited Bacterial RNA
7. RNAlater Preserves Bacterial Gene Expression Profiles for Array Analysis
8. Analysis of Bacterial Assemblage Genetic Diversity in Environmental Samples Using the DCode System
9. Monitoring Bacterial Genetic Diversity in a Freshwater Lake Using TTGE and DNA Sequence Analysis
10. Use of the DCode System to Detect the Food-Borne Bacterial Pathogen Listeria monocytogenes
11. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Bacterial RNA Isolation From Infected Eukaryotic Hosts

(Date:8/28/2015)... , Aug. 28, 2015 ... the addition of the "Global & USA BioSimilar ... The highly awaited US Biosimilar market will ... awaited filigrastim biologic, Zarxio. How will the global market ... author reveals that biosimilars could save the US economy ...
(Date:8/28/2015)... , August 28, 2015 According ... (Media, Sera, Cryogenic Storage Systems, Thawing Equipment, Alarms), by ... Medicine, Clinical Trials), End User - Global Forecast to ... expected to reach USD 3,731.03 Million by 2020 from ... of 11.65% between 2015 and 2020. ...
(Date:8/28/2015)... ... August 28, 2015 , ... ... two studies at the International Spine Intervention Society’ s 23rd Annual ... to August 1st, 2015. With a focus on evidence-based research and clinical application, ...
(Date:8/28/2015)... ... August 28, 2015 , ... Pittcon is pleased to ... a session on chemical spectroscopic imaging at JASIS 2015 (Japan Analytical & Scientific ... analytical and scientific instruments show which will be held at International Conference Hall ...
Breaking Biology Technology:Global & USA BioSimilar Market Analysis Report 2015-2021 2Global & USA BioSimilar Market Analysis Report 2015-2021 3Biopreservation Market Worth 3,731.03 Million USD by 2020 2Biopreservation Market Worth 3,731.03 Million USD by 2020 3Biopreservation Market Worth 3,731.03 Million USD by 2020 4Biopreservation Market Worth 3,731.03 Million USD by 2020 5Proove Biosciences Presents New Research Study at the International Spine Intervention Society’s Annual Scientific Meeting 2Proove Biosciences Presents New Research Study at the International Spine Intervention Society’s Annual Scientific Meeting 3Proove Biosciences Presents New Research Study at the International Spine Intervention Society’s Annual Scientific Meeting 4U.S. Symposium on Imaging Organized by Pittcon’s Program Chairman Chuck Gardner 2U.S. Symposium on Imaging Organized by Pittcon’s Program Chairman Chuck Gardner 3
... April 7, 2008 BioSpace, the world,s,leading online ... company,will host a Private Label Career Fair for ... will take place at the Hanover Marriott,in Whippany, ... (Logo: http://www.newscom.com/cgi-bin/prnh/20080222/CLF013LOGO ), Regeneron ...
... Microbia, Inc. today,announced that it has changed its ... long-lived tree that thrives in the harshest desert,environment, ... years. Known as a,"nurse tree," it provides a ... more organic matter for birds, reptiles,insects, animals, and ...
... 2008 BioSpace, the world,s,leading online bioscience job ... a Private Label Career Fair for Boehringer Ingelheim ... will be held at the Sheraton,Boston Hotel from ... http://www.newscom.com/cgi-bin/prnh/20080222/CLF013LOGO ), Boehringer Ingelheim is looking ...
Cached Biology Technology:BioSpace to Host Regeneron Pharmaceuticals Career Fair in Whippany, New Jersey 2Microbia Announces Name Change to Ironwood Pharmaceuticals 2Boehringer Ingelheim and BioSpace Search for Scientists at Career Fair in Boston, Massachusetts 2
(Date:8/28/2015)... -- According to a new market research ... (Pen & Paper Based, Hosted, Biometrics), Service, Application (Clinical ... Vertical and Region - Global Forecast to 2020", published ... from USD 2.4 Billion in 2015 to USD 7.5 ... (CAGR) of 25.6% from 2015 to 2020. ...
(Date:8/26/2015)... 26, 2015 The report "Multi-Factor Authentication ... Factor), Application (Travel & Immigration, Government, Banking, Defense, Commercial ... to 2020", published by MarketsandMarkets, Multi-Factor Authentication Market is ... at a CAGR of 17.7% between 2015 and 2020. ... igures spread through 169 P ...
(Date:8/25/2015)... CITY and BELLEVUE, Wash. ... leader in advanced robotic systems, announced today it will ... unmanned ground vehicle (UGV), at the National Tactical Officers ... The Guardian S is the first-ever ... culmination of years of research and in-field trials and ...
Breaking Biology News(10 mins):Cognitive Assessment and Training Market Worth 7.5 Billion USD by 2020 2Cognitive Assessment and Training Market Worth 7.5 Billion USD by 2020 3Cognitive Assessment and Training Market Worth 7.5 Billion USD by 2020 4Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 2Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 3Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 4Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 5Sarcos to Debut First-of-its-Kind Snake Robot 2Sarcos to Debut First-of-its-Kind Snake Robot 3
... China, a team of genome researchers has concluded in a ... large-scale gene re-sequencing. Their findings, which appear in the latest ... Sciences ( PNAS ), indicate that domesticated rice may ... ago in the Yangtze Valley of China. Previous research suggested ...
... itch adds to growing evidence that the chemical signals that ... make us wince in pain. The interactions between itch ... researcher Diana Bautista, an assistant professor of molecular and cell ... some nerve cells that respond only to itch and others ...
... after experts apply an extensive array of approaches. An ... rapidly determined the structure of protein molecules in several cases ... of the new method is reported May 1 in ... Dr. Frank DiMaio of the University of Washington (UW) in ...
Cached Biology News:Rice's origins point to China, genome researchers conclude 2Rice's origins point to China, genome researchers conclude 3Pain and itch connected down deep 2Pain and itch connected down deep 3Pain and itch connected down deep 4Several baffling puzzles in protein molecular structure solved with new method 2Several baffling puzzles in protein molecular structure solved with new method 3Several baffling puzzles in protein molecular structure solved with new method 4
Rabbit polyclonal to Warfarin ( Abpromise for all tested applications). Antigen: Chemical / Small Molecule conjugated to KLH...
Neutrophil cytosol factor 2 (NCF-2) (Neutrophil NADPH oxidase factor 2) (67 kDa neutrophil oxidase factor) (p67-phox) (NOXA2). [Source:Uniprot/SWISSPROT;Acc:P19878] Antigen: Recombinant Protein E...
Rabbit polyclonal to MA2 ( Abpromise for all tested applications). entrezGeneID: 10687 SwissProtID: Q9UL42...
... polyclonal to FANCM ( Abpromise for ... Synthetic peptide derived from within residues ... (Note: the amino acid sequence is ... ab27787 .) Entrez ...
Biology Products: