Product Focus: MicroRNA labeling with “Marker”-pCp

MicroRNAs (miRNAs) are short endogenous, non-coding RNAs that modulate gene expression through binding to messenger RNA. They may serve as cancer biomarkers, and various methods for their profiling and detection were reported[1].

Jena Bioscience’s approach relies on enzymatic attachment of a single “Marker”-pCp-nucleotide (pCp: Cytidine-3′,5′-bisphosphate) to the 3′-terminus of miRNA[2] (Fig. 1). Common markers are biotin and fluorescent dyes, and such labeled miRNAs can be isolated via streptavidin or directly applied to microarray expression profiling, respectively.

Products

Figure 1: A “Marker”-pCp is attached to the 3′-end of MicroRNA with T4 RNA ligase resulting in biotin- or fluorescence-labeled miRNA for further analysis.

Benefits

  • Simple direct labeling of sub-µg quantities of total RNA
  • No need for amplification or size fractionation
  • Little sequence bias and low detection limits
  • Direct miRNA measurement in complex samples

Selected References

  1. Yin et al. (2008) Profiling microRNA expression with microarrays. Trends in Biotechnology 26(2):70.
  2. Wang et al. (2007) Direct and sensitive miRNA profiling from low-input total RNA. RNA 13:151

Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

Product Focus: New Cas9 Case Study, Detection Kits and HR Vectors

Recently a new case study was published  from UC Davis’ Mouse Biology Program in which injection-ready Cas9 nickase mRNA and gRNA were used to flox a target gene and rapidly generate a mouse model with a conditional KO allele. SBI’s Cas9 SmartNickase mRNA and gRNA IVT Synthesis kits are highlighted.


 CRISPR/Cas9 SmartNuclease Systems

Genome Editing using guide RNAs
The recent discovery of the CRISPR/Cas9 complex has provided researchers an invaluable tool to target and modify any genomic sequence with high levels of efficacy and specificity. The system, consisting of an RNA-guided nuclease (Cas9) and guide RNA (gRNA) complementary to a target sequence, allows for sequence-specific cleavage of target loci across the genome.

  • Cas9 SmartNuclease All-in-one Plasmids – Wild-type and mutant CRISPR/Cas9 plasmids
  • Transfection and Injection-ready Cas9 mRNA and gRNA Synthesis Kit –  Ready-to-use synthetic Cas9 mRNA for in vivo applications
  • Cas9 Lentiviral Vectors and Pre-made Virus – Transduce primary cells and make stable Cas9 editing cell lines
  • Cas9 Detection Systems –  Cas9 Antibody and RT-PCR Primer Set Kits

Genome engineering has never been easier.

And yet, important questions remain for scientists wishing to take advantage of this technology in their research:

  • How do you make CRISPR/Cas9 technology work for your research?
  • With a number of available options for CRISPR/Cas9 reagents, which ones should you use?

 SBI’s PrecisionX™ Cas9 SmartNuclease™ system offers proven technology with unparalleled support and expertise to make your use of CRISPR/Cas9 easy and successful from the start.

Expertise

Application Notes:
SBI’s genome engineering specialists have created detailed protocols for first-time and experienced users of the CRISPR/Cas9 system, including details on gRNA and HR Targeting Vector design for the following applications:

SBI’s Provides

Cas9 Detection Kits

Accurately monitor the expression of the Cas9 editing enzyme using a validated Cas9 polyclonal antibody and qPCR/RT-PCR primer sets to detect and quantitate the Cas9 mRNA transcript expression levels.

New Positive and Negative Selection HR Vectors

SBI has released a new series of HR targeting vectors containing a PGK-hsvTK selection cassette that is located outside of the 3′ MCS homology arm cloning site. Use these vectors to screen against random integration events associated with plasmid transfection by using negative selection (hsvTK) to eliminate cells which have randomly integrated a donor plasmid instead of via homologous recombination at nuclease-mediated dsDNA breaks. When the constructs are randomly integrated into the genome, the PGK-hsvTK cassette is active and these cells will be sensitized to ganciclovir or fialuridine when added to the culture medium. Enrich for correctly-targeted cells and speed up your model cell line creation.

CRISPR/Cas9 Positive Control Kit

New to Cas9 Try our Cas9 Positive Control Kit (Cat# CAS605B-1), which includes an all-in-one Cas9/gRNA plasmid targeting the AAVS1 safe harbor locus, the EGIP 293T reporter cell line (developed at the NIH CRM), and an AAVS1/GFP HR donor to rescue EGFP expression in the EGIP control line. This kit now includes a T7 endonuclease, buffer and primers for Surveyor assay confirmation of target cutting as well, and a lower promotion price to make it easier to test CRISPR/Cas9 in any lab.


Where can I find more information about SBI?

Visit the manufacturer page at www.stratech.co.uk/SBI, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

Product Focus: Magic™ Mouse Adjuvant

Magic™ Mouse Adjuvant  (Code: CDN-A001-CRD) is a novel immunization adjuvant specifically designed for rapid production of high titers of antibodies in mice. The adjuvant contains immune-stimulatory CpG

DNA — short oligonucleotides that contain unmethylated cytosine-guanine dinucleotides within a certain base context. The mammalian immune system has evolved to recognize these sequences, which are found naturally in bacterial DNA, as a sign of infection. Exposure to CpG DNA results in very rapid and strong immune activation, and, when applied with an antigen, CpG DNA produces high titers of antigen-specific antibodies. Different CpG DNA sequences activate the immune systems of different animal species. Magic™ Mouse Adjuvant contains a proprietary DNA sequence that is specifically designed for immunization of mice.

Among all research adjuvants and vaccine adjuvants available in the market, Magic™ Mouse Adjuvant by far is the most efficient mouse adjuvant for all types of immunogens tested, in terms of its rapidness in raising immune responses and the antibody titers it produces. Of note, this adjuvant does not yield comparable titers in other species including other types of rodents.

Importantly, by avoiding making water-in-oil emulsions, heating, sonification, lyophilization, or homogenization in immunogen preparation, which destroys the native conformation of the immunogen, this adjuvant allows production of antibodies against conformational epitopes on native antigens.

KEY FEATURES

  1. Highly effective— rapid production of high titers of antibodies. Only 2 immunizations are necessary.
  2. Safe — non-toxic adjuvant with no adverse side effects to animals.
    Immunogen- and adjuvant-sparing —much less immunogen required than with conventional methods.
  3. Time-saving —strong immune responses can be reached within 35 days after immunization.
  4. Extremely valuable— in producing antibodies against conformational epitopes.

Where can I find more information about Creative Diagnostic?

Visit the manufacturer page at www.stratech.co.uk/creative-diagnostic, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

Product Focus: Culture cells in Exo-FBS™ media to avoid bovine exosome interference

  • FBS with bovine exosome-sized vesicles removed
  • Stripped of CD63 positive cow exosomes
  • No detectable cow microRNAs
  • Same cellular growth rates supported as standard FBS
  • Use the same as standard FBS (add 10% in DMEM or RPMI)

Exosomes isolated from cell culture media provide an exciting source for novel biomarker discovery and therapeutic potential. However, cells grown in FBS can have high levels of background from bovine serum, which typically contains an abundance of cow exosome vesicles. These cow exosomes can interfere with studies of exosomes secreted from cells of interest, for example, by contributing bovine microRNAs into a sample.

To improve research on tissue culture-derived exosomes, SBI has developed an exosome-depleted FBS growth supplement called Exo-FBS that has been stripped of bovine exosomes. Exo-FBS supports equivalent growth of many cell types in culture, is devoid of cow CD63 positive exosomes and does not have any measurable bovine microRNAs. Exo-FBS enables the study of cellular secreted exosomes in culture without the worry of contaminating cow exosomes. No ultracentrifugation is required. Exo-FBS is available as both a standard FBS supplement or heat-inactivated FBS media supplement (treated at 65°C for 15 minutes before bovine exosome removal).



Exo-FBS works with a broad range of cell types.
Sample Exo-FBS Product Citations:

Prostate Stromal Cells
Josson S, Gururajan M, Sung SY, Hu P, Shao C, Zhau HE, Liu C, Lichterman J, Duan P, Li Q, Rogatko A, Posadas EM, Haga CL, Chung LW. Stromal fibroblast-derived miR-409 promotes epithelial-to-mesenchymal transition and prostate tumorigenesis. Oncogene. 2014 Jul 28;0. doi: 10.1038/onc.2014.212.

Mesenchymal Stromal Cells
Xin H, Li Y, Liu Z, Wang X, Shang X, Cui Y, Gang Zhang Z, Chopp M. Mir-133b Promotes Neural Plasticity and Functional Recovery after Treatment of Stroke with Multipotent Mesenchymal Stromal Cells in Rats Via Transfer of Exosome-Enriched Extracellular Particles. Stem Cells. 2013 Apr 30. doi: 10.1002/stem.1409.

B-Cells
Momen-Heravi F, Bala S, Bukong T, Szabo G. Exosome-mediated delivery of functionally active miRNA-155 inhibitor to macrophages. Nanomedicine. 2014 Mar 29. pii: S1549-9634(14)00132-4.


Where can I find more information about SBI?

Visit the manufacturer page at www.stratech.co.uk/SBI, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

 

 

Can egg-based flu vaccines be replaced?

Expression of immunogenic influenza haemagglutinins in LEXSY


First example for expression of immunogenic influenza haemagglutinins in recombinant LEXSY

To beat the annual influenza threat, there is an urgent demand for fast and reliable pipelines for vaccine production. Although several novel recombinant platforms have gained some success (Schmidt 2013), the traditional egg-based production still remains the major source for seasonal flu vaccine.

Now, for the first time, Pion et al. (2014) demonstrated the expression of immunogenic influenza haemagglutinins (HA) in the eukaryotic protein expression system LEXSY.

Soluble HA proteins were secreted into the culture medium and affinity-purified with yields of up to 5 mg/L culture (Figure 1). Their immunogenicity was proven in a mouse model demonstrating suitability for antigen production. The overall process from cloning to purified recombinant HA was conducted in less than three months, a time frame in principle fully compatible with an influenza annual production campaign.

Figure 1: (A) 3D-Model of an influenza virus. HA = haemagglutinin antigen on the virus surface (from commons.wikimedia.org/wiki/File%3AFlu_und_legende_color_c.jpg). (B) SDS-PAGE of recombinant A/Vietnam/1194/04(H5N1) HA purified from LEXSY culture supernatants G: glycosylated, DG: deglycosylated HA proteins (from Pion et al. (2014)).


LEXSY products – all you need for cloning, expression strain selection, and cultivation

LEXSY Expression Kits
Convenient all-in-one expression kits allowing constitutive or inducible, intracellular or secretory expression of target proteins

LEXSY Strains & Cultivation Kits
The LEXSY expression hosts, as strains or complete ready-to-grow cultivation kits

LEXSY Cultivation Media
Complex and synthetic media for optimal growth of LEXSY strains

LEXSY Antibiotics
For efficient selection of recombinant strains

LEXSY Molecular Tools
Expression vectors and useful primers

PCR and Cloning enzymes
All restriction enzymes for the pLEXSY cloning sites, ligases, polymerases etc.


 References

  1. Schmidt (2013) Vaccines for pandemics. Nat. Biotech. 31:957.
  2.  Pion et al. (2014) Characterization and immunogenicity in mice of recombinant influenza haemagglutinins produced in Leishmania tarentolae. Vaccine 32:5570.

LEXSY – Eukaryotic protein expression in Leishmania tarentolae

The unicellular kinetoplast protozoan Leishmania tarentolae, isolated from the Moorish gecko Tarentola mauritanica, not pathogenic to mammalians (Biosafety level 1) – was turned into the protein-producing host of our eukaryotic protein expression system LEXSY:

  • eukaryotic host as easy to handle as E. coli: no specific labware, no cell biology equipment required
  • fully eukaryotic protein expression machinery with post-translational modifications, including glycosylation and disulfide bond formation
  • shuttle vectors: cloning in E. coli, expression in LEXSY host
  • constitutive or inducible, intracellular or secretory expression of target proteins
  • stable expression strains for constant protein production

 Downloadable Information

LEXSY Illustrated Information

Illustrations of LEXSY configurations, features and expression results are provided for download

LEXSY Manuals

Manuals with description of LEXSY configurations and detailled protocols for cultivation, cloning, transfection, selection and evaluation of expression strains are available for download

LEXSY Methods

Detailled protocols for cultivation, transfection, cryoconservation and handling of LEXSY are available for download

LEXSY Conference Presentations

Recent presentations of LEXSY technology on international conferences are available for download


Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

Selleck Chemicals News: New platform of high efficiency iPSC reprogramming

 

Induced pluripotent stem cells (iPSCs), which can be generated by reprogrammed somatic cells, provides a model for investigating cell developmental processes. Generation of iPSCs by somatic cells usually goes through the enforced expression of transcription factors including Oct4, Klf4, Sox2, and c-Myc. In a recent study published on Stem Cell Reports, Vidal et al. found the inhibition of transforming growth factor β (TGF-β) or activation of Wnt signaling, or both, can provide a high efficient iPSC reprogramming of different differentiated cells.

In the study, >80% mouse embryonic fibroblasts (MEFs) reprogrammed to iPSCs by inhibiting TGF-β together with activating Wnt signaling, in the presence of an enzymatic cofactor, ascorbic acid (AA). Reprogramming took only 1 week to reach this high efficiency. On the other hand, hepatoblasts and granulocyte-macrophage progenitors (GMPs), two progenitors, required only TGF-β inhibition or Wnt activation, respectively, to rapidly trigger pluripotency loci and to enter a pluripotent state. The reprogram processes were closed to 100% efficiency in less than a week. By further investigation, researchers suspected specific molecular features are responsible for the differences between fibroblasts and progenitor cells.

Collectively, the results reveal specific requirements of different somatic cells in process of iPSCs generation, as well as, provide a rapid and efficient platform for further investigating the mechanism of pluripotency induction.

 

Reference


Related Products

Cat.No. Product Name Information
S2745-SEL CHIR-98014 CHIR-98014 is a potent GSK-3α/β inhibitor with IC50 of 0.65 nM/0.58 nM, with the ability to distinguish GSK-3 from its closest homologs Cdc2 and ERK2.
S7566-SEL IM-12 IM-12 is a selective GSK-3β inhibitor with IC50 of 53 nM, and also enhances canonical Wnt signalling.

 


Where can I find more information about Selleck Chemicals?

Visit the manufacturer page at www.stratech.co.uk/selleck_chemicals, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

NEW Supplier: SynVivo – Cell-Based Microchip Platform

Stratech Scientific now supply, SynVivo (CFD Research Corporation) products.

SynVivo products are a physiological, cell-based microchip platform that provides a morphologically and biologically realistic microenvironment allowing real-time study of cellular behavior, drug delivery and drug discovery.
SynVivo’s patented technology for recreating microvasculature scale, shape, fluidics, and cellular constructs provides the most realistic in vitro testing platform for drug discovery, delivery, efficacy, and toxicity assays.

Static well plate and linear flow channel assays cannot differentiate between shapes of drug delivery vehicles, whereas SynVivo can. As particle shape becomes a more relevant area of study for drug delivery, SynVivo can provide new insights that can help optimize drug delivery.

SynVivo can be used to capture real-time distribution of drugs in the microvasculature. Stagnant and recirculation zones that affect overall drug delivery and distribution can be readily identified.SynVivo can be used to quantify drug particle distribution patterns through analysis of shear rate and adhesion.

In addtion, SynVivo can support the growth of endothelial cells, cancer cells and many other cell types for mimicking healthy and disease conditions. In addition, targeted drug delivery can be quantified and optimized using the in vivo morphology and fluidic conditions in SynVivo.

For further information on available products visit our product listing.

Below is a video illustrating how the system works.


Basic Research

Cellular behavior and microcirculatory dysfunction:

SynVivo chips can be used to study real-time cell-cell interactions for:

  • Inflammation (leukocyte-endothelium),
  • Cancer (tumor-endothelium),
  • Thrombosis (platelet-endothelium), and
  • Infection (microbial-endothelium)

under physiological fluid flow conditions representing healthy and pathological conditions.

Leukocytes rolling and adhering on endothelial cells Infection of brain neuronal cells Infection on neuronal cells

Where can I find more information about SynVivo?

Visit the manufacturer page at www.stratech.co.uk/synvivo, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


Product Focus: EZ-TAL Effector Assembly Kit – Genome editing at your fingertips

Complete TALE assembly kit

  • Simple 2-day protocol
  • Assemble specific TALE-Nucleases
  • Build your own transcription activators

The easiest way to make your own custom TALE Nucleases and Transciption Activators
Transcription activator-like effectors (TALEs) are a class of naturally occurring transcription effectors that recognize specific DNA sequences to modulate gene expression. First identified in the plant pathogen Xanthomonas, the modularity of the TALE DNA binding domain enables sequence-specific perturbation of gene function and offers broad applications in genetic and epigenetic studies.

The TALE binding domain is defined by a simple, yet elegant code of tandem repeat sequences called repeat-variable di-residues (RVDs) that allow for the predictable generation of customized TALEs that bind to any DNA sequence being targeted with high levels of specificity. Compared to other gene targeting technologies such as zinc-finger nucleases (ZFNs) and meganucleases, TALEs offer the researcher more latitude in selecting potential DNA targets, ease in design, and provides levels of specificity on par or better than the aforementioned technologies.


EZ-TAL Kit Components and Assembly Workflow

Everything you need to make your own TALEs
Each EZ-TAL Kit contains the following components with enough material to perform a minimum of 6 and a maximum of 12 TALE assemblies, depending on the target sequence redundancy. The products are supplied in two boxes. Box 1 contains the entire TALE monomer library to built the multimers. Box 2 has all of the enzymes, buffers, substrates and colony PCR screening primers. Choose a genomic sequence target region and use the online software: TAL Effector Nucleotide Targeter website. Once your target sequences are selected, use the EZ-TAL ready-made monomer library to begin building your custom TAL Effector in just 2 days. The first step of the do-it-yourself protocol is to assembled monomers into multimers using a procedure based on restriction, ligation and amplification. Multimers 1 and 2 are designed to be hexamers, but the length of multimer 3 can vary to allow variations in the final length, such as 14-19 bp binding sequences. The correctly assembled circular multimers were subsequently amplified by PCR. On day 2, gel-purified multimers are assembled into a vector of choice, using a second restriction-ligation-based procedure, followed by bacterial transformation. Colony PCR is then performed for confirmation of insert size; typically, 40-90% of colonies displayed correct insert size. We recommend to use two colonies of correct insert size for sequence confirmation; typically 80-90% of sequences revealed correct assembly. The last step is to test your TAL Effectors for activity, the surveyor nuclease assay can be used to determine cutting efficiency at the desired genomic location. A schematic of the entire work flow is shown below.


Products

 

EZ-TAL Nuclease and Transcription factor Assembly Kits

EZ-TAL Assembly Kit-CMV-TALEN 1 Kit GE100A-1
EZ-TAL Assembly Kit-EF1a-TALEN 1 Kit GE120A-1
EZ-TAL Assembly Kit-MSCV-TALEN 1 Kit GE140A-1
EZ-TAL Assembly Kit-CMV-TALE-TF 1 Kit GE500A-1
EZ-TAL Assembly Kit-EF1a-TALE-TF 1 Kit GE520A-1
EZ-TAL Assembly Kit-MSCV-TALE-TF 1 Kit GE540A-1
EZ-TAL End-Monomer Kit (Contains end-monomers at positions 12, 13, 14, 15, 16, 17). Compatible with GE1xx-5xxA-1 (required) to generate TALEs with various lengths of 14 to 19-mers. 1 Kit provides a minimum of 6 TALEN assemblies. 1 Kit GE900A-1
EZ-TAL Multimer Amplification Primer Mix (10uM) 50 ul GE590PR-1
EZ-TAL Colony PCR Primer Mix (10uM) 50 ul GE595PR-1

 

For TALE-Nuclease assembly, SBI recommends Catalog # GE100A-1, as the CMV promoter is generally the strongest and most widely used option in the literature for TALEN applications.

 

TAL Effector Services

TALE Nuclease pair for sequence-specific??gene editing 10 ul plasmid DNA CS810TN-1
Artificial gene activator with a custom TALE DNA binding domain 10 ul plasmid DNA CS820TA-1
Artificial gene inhibitor with a custom TALE DNA binding domain 10 ul plasmid DNA CS820TR-1
A dual-reporter (GFP and luciferase) for monitoring TA/TR functionality 10 ul plasmid DNA CS830DR-1

Online Resources


Product Citations

  1. Uhde-Stone C, Sarkar N, Antes T, Otoc N, Kim Y, Jiang YJ, Lu B. A TALEN-based strategy for efficient bi-allelic miRNA ablation in human cells. RNA. 2014 Apr 9. [Epub ahead of print].
  2. Uhde-Stone C, Gor N, Chin T, Huang J, Lu B. A do-it-yourself protocol for simple transcription activator-like effector assembly. Biol Proced Online. 2013 Jan 14;15(1):3. doi: 10.1186/1480-9222-15-3.

Technical References

  1. Bedell, V. M., Y. Wang, J. M. Campbell, T. L. Poshusta, C. G. Starker, R. G. Krug Ii, W. Tan, S. G. Penheiter, A. C. Ma, A. Y. Leung, S. C. Fahrenkrug, D. F. Carlson, D. F. Voytas, K. J. Clark, J. J. Essner & S. C. Ekker (2012) In vivo genome editing using a high-efficiency TALEN system. Nature, doi: 10.1038/nature11537.
  2. Boch, J., H. Scholze, S. Schornack, A. Landgraf, S. Hahn, S. Kay, T. Lahaye, A. Nickstadt & U. Bonas (2009) Breaking the code of DNA binding specificity of TAL-type III effectors. Science, 326, 1509-12.
  3. Clark, K. J., D. F. Voytas & S. C. Ekker (2011) A TALE of two nucleases: gene targeting for the masses? Zebrafish, 8, 147-9. Doyle, E. L., N. J. Booher, D. S. Standage, D. F. Voytas, V. P. Brendel, J. K. Vandyk & A. J. Bogdanove (2012) TAL Effector-Nucleotide Targeter (TALE-NT) 2.0: tools for TAL effector design and target prediction. Nucleic Acids Res, 40, W117-22.
  4. Hockemeyer, D., H. Wang, S. Kiani, C. S. Lai, Q. Gao, J. P. Cassady, G. J. Cost, L. Zhang, Y. Santiago, J. C. Miller, B. Zeitler, J. M. Cherone, X. Meng, S. J. Hinkley, E. J. Rebar, P. D. Gregory, F. D. Urnov & R. Jaenisch (2011) Genetic engineering of human pluripotent cells using TALE nucleases. Nat Biotechnol, 29, 731-4.
  5. Huang, P., Z. Zhu, S. Lin & B. Zhang (2012) Reverse genetic approaches in zebrafish. J Genet Genomics, 39, 421-33. Lei, Y., X. Guo, Y. Liu, Y. Cao, Y. Deng, X. Chen, C. H. Cheng, I. B. Dawid, Y. Chen & H. Zhao (2012) Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs). Proc Natl Acad Sci U S A, PMID: 23045671.
  6. Li, L., M. J. Piatek, A. Atef, A. Piatek, A. Wibowo, X. Fang, J. S. Sabir, J. K. Zhu & M. M. Mahfouz (2012a) Rapid and highly efficient construction of TALE-based transcriptional regulators and nucleases for genome modification. Plant Mol Biol, 78, 407-16.
  7. Li, T., S. Huang, X. Zhao, D. A. Wright, S. Carpenter, M. H. Spalding, D. P. Weeks & B. Yang (2011) Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes. Nucleic Acids Res, 39, 6315-25. Li, T., B. Liu, M. H. Spalding, D. P. Weeks & B. Yang (2012b) High-efficiency TALEN-based gene editing produces disease-resistant rice. Nat Biotechnol, 30, 390-2.
  8. Liu, J., C. Li, Z. Yu, P. Huang, H. Wu, C. Wei, N. Zhu, Y. Shen, Y. Chen, B. Zhang, W. M. Deng & R. Jiao (2012) Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy. J Genet Genomics, 39, 209-15.
  9. Ma, S., S. Zhang, F. Wang, Y. Liu, Y. Liu, H. Xu, C. Liu, Y. Lin, P. Zhao & Q. Xia (2012) Highly Efficient and Specific Genome Editing in Silkworm Using Custom TALENs. PLoS One, 7, e45035.
  10. Mahfouz, M. M. & L. Li (2011) TALE nucleases and next generation GM crops. GM Crops, 2, 99-103.
  11. Mahfouz, M. M., L. Li, M. Piatek, X. Fang, H. Mansour, D. K. Bangarusamy & J. K. Zhu (2012) Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein. Plant Mol Biol, 78, 311-21.
  12. Moore, F. E., D. Reyon, J. D. Sander, S. A. Martinez, J. S. Blackburn, C. Khayter, C. L. Ramirez, J. K. Joung & D. M. Langenau (2012) Improved somatic mutagenesis in zebrafish using transcription activator-like effector nucleases (TALENs). PLoS One, 7, e37877.
  13. Mussolino, C. & T. Cathomen (2012) TALE nucleases: tailored genome engineering made easy. Curr Opin Biotechnol, 23, 644-50. Perez-Pinera, P., D. G. Ousterout & C. A. Gersbach (2012) Advances in targeted genome editing. Curr Opin Chem Biol, 16, 268-77.
  14. Reyon, D., S. Q. Tsai, C. Khayter, J. A. Foden, J. D. Sander & J. K. Joung (2012) FLASH assembly of TALENs for high-throughput genome editing. Nat Biotechnol. 30, 460-5.
  15. Uhde-Stone, C., J. Huang & B. Lu (2012) A robust dual reporter system to visualize and quantify gene expression mediated by transcription activator-like effectors. Biol Proced Online, 14, 8.

Where can I find more information about SBI?

Visit the manufacturer page at www.stratech.co.uk/SBI, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.

 


Study Exosomes from Culture Media without FBS Contamination

Unless you are studying Cow exosomes, use Exo-FBS when isolating exosomes from cell culture media.

Exosomes isolated from cell culture media provide an exciting source for novel biomarker discovery and therapeutic potential. However, cells grown in FBS can have high levels of background from bovine serum, which typically contains an abundance of cow exosome vesicles. These cow exosomes can interfere with studies of exosomes secreted from cells of interest, for example, by contributing bovine microRNAs into a sample.

To improve research on tissue culture-derived exosomes, SBI has developed an exosome-depleted FBS growth supplement called Exo-FBS that has been stripped of bovine exosomes. Exo-FBS supports equivalent growth of many cell types in culture, is devoid of cow CD63 positive exosomes and does not have any measurable bovine microRNAs. Exo-FBS enables the study of cellular secreted exosomes in culture without the worry of contaminating cow exosomes. No ultracentrifugation is required. Exo-FBS is available as both a standard FBS supplement or heat-inactivated FBS media supplement (treated at 65°C for 15 minutes before bovine exosome removal).

HIGHLIGHTS

  • FBS with bovine exosome-sized vesicles removed
  • Stripped of CD63 positive cow exosomes
  • No detectable cow microRNAs
  • Same cellular growth rates supported as standard FBS
  • Use the same as standard FBS (add 10% in DMEM or RPMI)

Exo-FBS works with a broad range of cell types.


Sample Exo-FBS Product Citations:

Prostate Stromal Cells
Josson S, Gururajan M, Sung SY, Hu P, Shao C, Zhau HE, Liu C, Lichterman J, Duan P, Li Q, Rogatko A, Posadas EM, Haga CL, Chung LW. Stromal fibroblast-derived miR-409 promotes epithelial-to-mesenchymal transition and prostate tumorigenesis. Oncogene. 2014 Jul 28;0. doi: 10.1038/onc.2014.212.

Mesenchymal Stromal Cells
Xin H, Li Y, Liu Z, Wang X, Shang X, Cui Y, Gang Zhang Z, Chopp M. Mir-133b Promotes Neural Plasticity and Functional Recovery after Treatment of Stroke with Multipotent Mesenchymal Stromal Cells in Rats Via Transfer of Exosome-Enriched Extracellular Particles. Stem Cells. 2013 Apr 30. doi: 10.1002/stem.1409.

B-Cells
Momen-Heravi F, Bala S, Bukong T, Szabo G. Exosome-mediated delivery of functionally active miRNA-155 inhibitor to macrophages. Nanomedicine. 2014 Mar 29. pii: S1549-9634(14)00132-4.


Where can I find more information about SBI?

Visit the manufacturer page at www.stratech.co.uk/SBI, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


 

 

Product Focus: dNTPs & dNTP Pre-Mixes

Premium Quality dNTPs – at prices you can’t ignore…!

The synthesis of a complementary DNA/cDNA strand by enzymatic polymerization of deoxynucleotide triphosphates (dNTPs) is the basic principle of DNA amplification, sequencing and cDNA synthesis techniques such as PCR, real-time PCR and reverse transcription. For these, amplification sensitivity, product yield and reproducibility are strongly dependent on the quality of the reagents used.

Jena Bioscience’s enzymatic dNTP manufacturing process ensures superior quality for deoxynucleotide triphosphates: All their dNTPs are ultrapure (> 99 % by HPLC) and functionally tested by a set very sensitive enzymatic reactions (see below dNTP Guide and dNTP specifications).

Jena Bioscience dNTP Specifications


dNTP Mix / 10 mM

Premix of 10 mM dATP, dCTP, dGTP and dTTP

Product Cat. No. Amount
S pack NU-1006S-JEN 200 µl
L pack NU-1006L-JEN 1 ml

dNTP Mix / 25 mM

Premix of 25 mM dATP, dCTP, dGTP and dTTP

Product Cat. No. Amount
S pack NU-1023S-JEN 200 µl
L pack NU-1023L-JEN 1 ml

dNTP Mix incl. dUTP / 10 mM

Premix of 10 mM dATP, dCTP, dGTP and 20 mM dUTP

Product Cat. No. Amount
S pack NU-1020S-JEN 200 µl
L pack NU-1020L-JEN 1 ml

dNTP Mix GCamplifier / 10 mM

Modified dNTP Mix for amplification of GC-rich sequences

Product Cat. No. Amount
Kit PCR-257-JEN 100 µl

dNTP PCR Mix GCamplifier is specially designed for amplification of GC-rich DNA templates. The kit contains a modified dGTP analog that significantly reduces the stability of GC-rich sequences thereby enhancing PCR efficiency. The dNTP PCR Mix GCamplifier is used instead of a standard dNTP mix.


Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.