Frequently Asked Questions
General
RNAi Combination Toolbox™
Services
Do you provide synthetic siRNA duplexes?
No. We are specialized in RNAi expression vector design. Studies have shown that shRNA expressed from
plasmid vector work better than synthetic siRNA duplexes. In addition, vector-based RNAi has several advantages over synthetic
siRNA duplexes (see below).
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What are the advantages to use plasmid-based RNAi expression vectors?
Plasmid-based RNAi expression vectors have several advantages over the synthetic siRNA duplexes:
First of all, shRNAs/miRNAs expressed from a plasmid vector work better and last longer than synthetic siRNA duplexes.
Secondly, DNA vectors are stable and easy to handle. Transfection with plasmid DNA on eukaryotic cells is a familiar technique
in many research labs. Moreover, a vector carrying selection marker allows you to establish stable cell lines to observe the
long-term effects of gene silencing. Finally, a plasmid-based RNAi expression vector is cost-effective. Preparation of an RNAi
expression vector construct is a one-time investment. Unlike synthetic siRNA duplexes, which have to be repeatedly ordered,
plasmid DNA can be easily maintained and amplified at low costs.
Through innovation, LBL scientists have added special features (see below) onto plasmid-based RNAi
expression vectors, which allow you to perform RNAi experiments far beyond the capacity of what synthetic siRNA duplexes can do.
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I have already been using other RNAi vectors. Do I have to switch to your system?
RNAi Combination Toolbox™ vectors have additonal features that you can not find from other
RNAi expression vectors (see below). Switching to our system would allow you to do more kinds of RNAi
experiments, and greatly facilitate your gene function analysis.
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What if I still have questions that are not found in your list?
If you want to know more about our products, services or
RNAi technology in general, send your questions to our Technical Support Team by e-mail at:
support@lanleys.com
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What is the difference between your RNAi expression vectors and others?
RNAi Combination Toolbox™ is a plasmid-based convertible gene silencing system developed by
LBL scientists, which is the first of this kind in RNAi technology field. It contains sets of specially designed vectors, which
can maximize your strength to use RNAi technology in a cost-effective manner.
Among RNAi Combination Toolbox™ vectors, DNA hairpin can be moved from one vector to another by simple steps of DNA
subcloning to meet your research needs. Our vectors have common features of all other RNAi expression vectors, such as
in vitro transcription, expression of shRNA/miRNA in the cells and establishment of stable cell lines. In addition, our
vectors allow you to generate multiple RNAi expression cassettes or co-express RNAi and rescue cDNA from a single vector, which
constitute unique features that you can not find on any other vectors.
LBL scientists are in a continuing effort to develop more RNAi expression vectors based on your need in RNAi experiments. It
is our hope that RNAi technology will soon become a more affordable and easily adapted molecular tool in all research labs.
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Do I have to buy all the vectors from your RNAi Combination Toolbox™?
No. RNAi Combination Toolbox™ contains sets of RNAi expression vectors for various research
purposes. In most cases, one vector product will be good enough for you to perform one or more types of RNAi experiments.
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Do you have a vector that can do everything?
No. We don't have a magic wand to make such a super vector that can do everything for you.
However, RNAi Combination Toolbox™ contains vectors that are often able to carry out
multiple tasks to meet your research needs. Check technical information of each vector for details.
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I already have a validated siRNA. Can I clone it into your vectors?
Yes. You just need to convert your siRNA sequence to a DNA hairpin and clone it into one of our RNAi expression vectors.
Learn more on how to Design your DNA Hairpin for RNAi Combination Toolbox™.
Notably, DNA hairpin from converted siRNA may not always work the same as original siRNA duplexes for various reasons. If
you have any question or encounter any difficulty, write to our Technical Support Team by e-mail at:
support@lanleys.com
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How can I design my DNA hairpin and subclone it into your vectors?
Please read a detail description on how to Design your DNA Hairpin for RNAi Combination Toolbox™.
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What kind of transfection reagents should I use for your plasmid vectors?
Any transfection reagent that works for you with any other DNA plasmids will be OK. Nothing is special for transfecting
RNAi Combination Toolbox™ vectors.
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What is the difference between your multiple expression cassettes and chained shRNA/miRNA hairpins from other suppliers?
Some vectors from other suppliers allow you to generate chained shRNA/miRNA hairpins, which are transcribed by a single
Pol III/Pol II promoter. However, our RNAi expression vectors allow you to generate multiple expression cassettes, where each
cassette contains an RNAi expression unit. Therefore, every shRNA/miRNA hairpin has its own promoter, which can maximize its
expression in the cells.
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I can not find Csp45I on your restriction maps. Why?
AsuII works the same as Csp45I.
Check the list of Isoschizomers on the multiple cloning sites of our vectors. Please note
that suppliers may have different isoschizomers in their product lists.
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Can I use Swapping/Docking sites other than those provided on the technical information sheet?
Yes. The Swapping and Docking sites provided on technical information sheet are designed for your convenience to perform DNA
subcloning. It is absolutely of your choice to use any other restriction sites, which can do the same job.
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How many RNAi expression cassettes can be subcloned into a single vector?
We haven't determined the absolute maximum number of expression cassettes that can be subcloned into a single vector.
Typically a plasmid vector ranging from 2.0 kb to 4.0 kb in size can uptake an insert over 5 kb or more in length, which is
about the size of 12 RNAi expression cassettes (~ 400 bp each).
Notably, RNAi is a biological process and saturable in the presence of excessive amount of double-stranded RNAs. Normally 3 - 5
genes can be targeted simultaneously without significant reduction in the gene silencing capacity of each cassette.
Therefore, it is the effectiveness of RNAi machinery in the cells, not the size of the plasmid, that becomes the limiting factor
when you want to silence multiple gene targets from a single vector.
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Can I do co-transfection with several RNAi expression vectors?
Yes. RNAi expression vectors from RNAi Combination Toolbox™ can be transfected as
conventional DNA plasmids. Co-transfection will work the same way for these vectors.
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Can I use restriction enzymes other than PstI for diagnostic analysis?
Yes. PstI is recommended only when you are planning to subclone your DNA hairpin into vectors for
in vitro transcription. Otherwise, it is of your choice to determine the use of
restriction enzymes for diagnostic analysis.
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Why do you charge fees on RNAi target selection service?
Many simplified version of RNAi target selection programs are available on the web to let you design RNAi targets by yourself
with minimal to some levels of flexibility in design parameters, which usually result in moderate effects.
RNAlysis™ RNAi Target Selection Protocol is developed by LBL scientists to achieve the best
gene silencing result, while keep off-target effect to the least. In addition, we provide the most flexible customized
RNAi target selection service, which is unique and involves substantial amount of manpower and intellectual input. For example,
our experienced scientists can sort out selective targets among gene isoforms or family genes depending on your research
interests, which can not be simply carried out by any computer program. Please note that, this service is free for
customers, who purchase additional services and/or products from LBL.
It is our mission to provide you the most advanced RNAi technology and let you focus on your research goals.
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Will you be able to provide final plasmid DNA construct if I have nothing but a paper sequence and its design?
Yes, we will be able to provide you desired plasmid DNA construct. If you have more materials to start with, it will cost less.
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Do you have a complete gene targeting and transgenic service?
No. We have expertise to prepare and handle extra large vectors, which are typically 30 - 50 kb in total, for gene targeting
applications. We provide strategy design, isolation and characterization of genomic clone, mapping and construction services of
targeting vector for your gene knock-in or knock-out studies.
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If I have cDNAs in other expression vectors, can you do the yeast-two hybrid for me?
Yes. We will subclone your cDNAs into yeast-two hybrid vectors, and then do the test for you.
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How is your yeast-two hybrid service different from others?
Yeast-two hybrid has been proven to be an efficient molecular and cell biology technique for protein interaction analysis, which
is often required in many studies. In LBL, we have developed our own specialities to master this useful tool.
We provide high quality services with very competitive price. For example, after library screening our customers will receive
the full length sequence of positive clones, while other service providers may only give you partial, usually the 5'-end sequence
information. Having the most accurate identity of positive clones will ultimately save your time on mapping the interaction sites.
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My project is confidential. How do you handle this work?
We can sign a confidentiality agreement upon request.
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