Based on a novel variety of Magnetic Nanoparticles, ElementZero have just launched a unique technology, allowing specific capture and enrichment of a broad variety of viral RNA (SARS-CoV-2, HIV, Ebola….), or any of your specific RNA or DNA targets. Discover these new products, now available at in Europe through tebu-bio.
tebu-bio is pleased to announce their sponsorship of the 1st Course on “Post-transcriptional gene regulation: mechanisms at the heart of networks” (Institut Curie, March 23-27, 2015) that will be training 40 PhD students and postdoctoral fellows from 10 countries. Post-transcriptional gene regulation refers to every level of gene-expression control in a cell that occurs between transcription and the resulting protein levels. Specialists in this field study such things as polyadenylation, miRNAs, RNA binding proteins, signal transduction, and (pre)mRNA splicing.
If you don’t buy luxury brand clothes, why are you still buying luxury brand molecular biology kits?
Let me guess… you might use those blue and red boxed Qiagen® kits for DNA and RNA purification, maybe TRIzol® if you’re a real rebel. You probably also use a SuperScript® RT Kit for reverse transcription and you can’t keep track of which company most recently bought Invitrogen® to control that logo that you’re loyal to.
Maybe it used to be the case that these were the only companies making these kits, so you really had no choice. Now, many of these products are essentially commodities, but you’re still used to purchasing the “first to market” brand because it works and it would probably cause trouble for your lab to change brands. The perceived “switching costs” are large part of the reason for this persistent brand loyalty, and part of the reason why ThermoFisher was willing to pay $13.6 billion for the Life Technologies brand in 2013. There is an entire arm of corporate finance devoted to assigning the value to a brand. Essentially, it tries to estimate how much more customers are willing to pay for a commodity item just because of the company logo on the side of the package.
Here, we examine some alternatives to 2 of the biggest molecular biology mega-brands that seem to be able to keep their loyal customers by placing their familiar brand name on a relatively low value item:
1 – Alternatives to Lipofectamine® Reagents
There’s the one with the blue cap, the one with the red cap, and the one with the green cap, and most people don’t really understand the difference, but cell biologists seem to love these transfection reagents and are willing to pay a heavy premium for them.
Compared to the prices announced in 2015 on the Life Tech website for the 1.5 mL size Lipofectamine® 2000, similar products from competitors are available for about 1/4 the price:
– Enhanced conjugate with liposome for maximal efficiency
– Best for lentivirus, antibody and protein production
GenMute™ siRNA 1 ml
– Exceptionally good for siRNA transfection
– Excellent for DNA/siRNA co-transfection
LipoJet™ DNA 1 ml
– Fluorinated cationic lipids
– Best transfection efficiency with least toxicity
Validation data for Lipojet™, for example, suggest that it is superior to many other commercial transfection reagents including “L2K”::
Given the vast selection of transfection reagents available today, the best option is usually to contact a technical specialist (at tebu-bio for example) for advice on the most suitable ones for your research.
2 – Alternatives to Qiagen® and TRIzol®
Whether you’re using the DNeasy® and RNeasy® kits or TRIzol® products for DNA and RNA extraction, chances are you just keep buying the same kit you learned to use when you were a student. Sure, there are companies making less expensive column-based kits, and there are even protocols online explaining how to regenerate the used columns and make your own buffers. You can also find generic RNAzol RT RNA Isolation Reagent.
The advances we really like, however, are the purification systems that work without columns and without dangerous solvents. Here are a few of our favourites:
QuickExtract™ DNA Extraction Solution The fastest, least expensive option when you just need to purify DNA for PCR.
QuickExtract™ RNA Extraction Kit Available in two sizes: the extremely competitively priced 5mL kit contains sufficient material to perform 50 RNA extractions, while the 50mL kit contains sufficient material to perform 500 RNA extractions.
MasterPure™ DNA and RNA Purification Kits These are for the times when you need the highest quality nucleic acid for next generation sequencing, qPCR, or PCR. Produced by the RNA biology experts at Epicentre (an Illumina company), MasterPure™ Kits are available for nearly every application and source including mammalian cells, yeast, blood, Gram Positive bacteria, and Plant Leaves.
As is the case with transfection reagents, choosing a new nucleic acid purification kit is not always easy, so feel free to contact a technical specialist at tebu-bio for advice.
To get in touch, just leave your comments or questions below.
DNeasy® and RNeasy®, and Qiagen® are registered trademarks of the Qiagen group. TRIzol® is a registered trademark of Molecular Research Center, Inc. Invitrogen® and SuperScript® are registered trademarks of Invitrogen. Lipofectamine® is a registered trademark of Life Technologies, Inc.
In vitro transcription has been a common protocol in RNA biology laboratories wishing to work directly with mRNA molecules to study phenomena such as mRNA translation. Commercially available kits have greatly facilitated the capping and polyadenylation and in vitro transcription of RNAs, but now there is another option: ordering chemically synthesized mRNAs.
In vitro transcription kits such as the T7-FlashScribe™ Transcription Kit allow maximum RNA yields in 30 minutes. Subsequent processing of primary transcripts involves addition of the 5′ cap and 3′ poly(A) tail.
Some of the most popular kits for mRNA processing include the ScriptCap™ m7G Capping System for capping and the A-Plus™ Poly(A) Polymerase Tailing Kit for tailing. Researchers wishing to optimize protein expression use chemically-modified mRNAs, such as those carrying an anti-reverse cap analog ARCA at the 5′ end. Again, kits such as the MessageMAX™ T7 ARCA-Capped Message Transcription Kit have made in vitro transcription of ARCA capped mRNAs routine in laboratories. When a standard m7 GTP cap is added to mRNAs in vitro, only about 1/2 of the cap is added in the correct orientation. The ARCA cap is modified with a methyl group to prevent capping in the “incorrect” orientation, thus resulting in a higher percentage of efficiently translated mRNA.
Depending on the application, researchers are seeing advantages of using other chemical modifications as well. Pseudouridine-5′-Triphosphate, a naturally occurring base, is used to decrease nuclease activity and TLR activation, and modified cytidine base (5-Methylcytidine-5′-Triphosphate) is used for similar reasons. Warren et al. 2010 (doi: 10.1016/j.stem.2010.08.012), for example, found that mRNAs carrying an ARCA cap and Pseudo-uridine and methyl-cytidine substitutions are very efficient for reprogramming of many human cell types and fail to activate the toll-like receptor innate immune pathways.
Similarly, the Immune Stimulation Transcription Nucleotide Set is available for those who want to purchase ARCA cap, Pseudouridine-5′-Triphosphate, and 5-Methylcytidine-5′-Triphosphate. Another modification gaining popularity is the modified uridine (2-Thiouridine-5′-Triphosphate), however this popularity may be a result only of intellectual property limiting the commercial use of Pseudo-uridine.
Commercially-available capped and polyadenylated mRNAs modified with pseudouridine and 5-methylcytidine include those encoding:
|EGFP mRNA (5meC, Ψ)||+||+||996|
|Oct4 mRNA (5meC, Ψ)||+||+||1,359|
|Klf4 mRNA (5meC, Ψ)||+||+||1,688|
|SOX2 mRNA (5meC, Ψ)||+||+||1,230|
|c-Myc mRNA (5meC, Ψ)||+||+||1,596|
|Lin28 mRNA (5meC, Ψ)||+||+||906|
|FLuc mRNA (5meC, Ψ)||+||+||1‚929|
|NLS-Cre mRNA (5meC, Ψ)||+||+||1‚350|
|β-gal mRNA (5meC, Ψ)||+||+||3,336|
|Factor IX mRNA (5meC, Ψ)||+||+||1,662|
|hAAT mRNA (5meC, Ψ)||+||+||1,530|
|mCherry mRNA (5meC, Ψ)||+||+||996|
|Eira CFP mRNA (5meC, Ψ)||+||+||978|
|Blaze YFP mRNA (5meC, Ψ)||+||+||987|
|Cas9 Nickase mRNA (5meC, Ψ)||+||+||4,341|
|EPO mRNA (5meC, Ψ)||+||+||858|
|CD8 mRNA (5meC, Ψ)||+||+||coming soon|
|NGFR mRNA (5meC, Ψ)||+||+||coming soon|
|Guassia Luciferase mRNA (5meC, Ψ)||+||+||834|
|Renilla Luciferase mRNA (5meC, Ψ)||+||+||1,212|
|Cas9 mRNA (5meC, Ψ)||+||+||4,509|
|EGFP mRNA (5meC)||+||996|
|FLuc mRNA (5meC)||+||1‚929|
|OVA mRNA (5meC, Ψ)||+||+||1,437|
|Cas9 mRNA (Ψ)||+||4,509|
|Cyanine 5 FLuc mRNA (5meC, Ψ)||+||+||1‚929|
|Cyanine 5 EGFP mRNA (5meC, Ψ)||+||+||996|
Clearly, many of these purified mRNAs encoding fluorescent proteins, luciferase, or other reporter proteins are intended as controls for users setting up assays on which disease-relevant mRNAs will be tested. Once the assays are established, users now have the choice to produce their mRNAs of interest themselves using the in vitro transcription, capping, and polyadenylation kits described above or to order high quality mRNAs produced by expert chemists. Chemically synthesized mRNAs can be synthesized to contain nearly any sequence and chemical modification desired and gram quantity yields are possible. For particularly long mRNAs (up to multiple kilobases), in vitro transcription steps may still be required, but experts at TriLink Biotechnologies are able to design custom strategies to optimize yield even for the most complicated custom mRNA production requirements.
European scientists interested in learning more about out-sourcing mRNA production are encouraged to contact the local TriLink Biotechnologies distributor, tebu-bio.