How do siPOOLs eliminate off-target effects?
An siPOOL, is a complex mixture of 30 selected siRNAs directed against the target gene, which are transiently transfected into target cells using well established protocols and reagents. By using 30 different siRNAs against the same target gene, off-target effects of each siRNA are diluted below experimental relevance. At the same time all 30 siRNAs in the pool ensure an efficient gene silencing.
How much do I get if I order a siPOOL?
For standard set up siPOOLs are available at 5 nmol or 10 nmol per target gene. In addition positive and negative control pools are available. Please contact us if you need to order different amounts at firstname.lastname@example.org
How long does it take from placing an order to delivery?
We do need approximately 2-3 weeks from the date you place your order to delivery. After you place an order we will send you an order confirmation stating also the approximately delivery time. In many cases we can shorten the delivery time. However, the synthesis of siPOOLs involves research, therefore delivery dates for siPOOLs and validation data cannot be guaranteed. Galen Laboratory Supplies will give notification in due time if the delivery will be delayed.
What concentration of siPOOLs shall I use for transfection?
siPOOLs are very efficient in gene knock-down. In standard cell lines a total concentration of 1 – 3 nM is sufficient. A detailed transfection protocol for standard applications in 24 or 96 well plates is provided with every siPOOL kit.
Are transfection conditions using siPOOLs different from transfection conditions using other siRNA reagents?
No, you may apply exactly the same standard transfection conditions for siPOOLs as for other siRNA reagents. For most cell lines, lipofection with Lipofectamine works fine following the instructions of the provider.
Which transfection reagent do you recommend?
There is a broad range of transfection reagents commerically available. For many common cell lines Lipofectamine works very well. However, transfection of cell types like primary macrophages or non-adherent cells can be tricky. If you work with such cells or are not sure about optimizing your transfection protocol, please conduct us, we are happy to assist in the optimization of your experimental set up.
What is the difference between your product and the other commercially available pools of siRNAs?
Other commercially available siRNA pools are either low complexity pools, or stochastic pools. In contrast siPOOLs are the first available high complex pools - 30 siRNAs - of carefully selected and defined siRNAs.
How long will the duration of silencing be?
The duration of RNAi effects strongly depends on the cell line and
target-gene. However in general one can assume that the expression of
the target gene will be silenced (= down-regulated) for several days.
Working in standard cell lines like A549, MCF7, Hela etc. most genes
should stay down for more than 48h, probably even more than 72h.
What is your negative control siPOOL?
The standard negative control siPOOL is a pool of 30 siRNAs which do not interact with human, mouse and rat genes and can therefore be used in those species for standard experiments. If you are ordering a siPOOL targeting genes in other species, we will provide a suitable negative control siPOOL if required.
What is your positive control siPOOL and what is the read-out?
The positive control siPOOL consists of 30 selected siRNAs targeting the PLK-1 gene. Knock-down of PLK1 expression does inhibit cell growth and induces pro-apoptotic pathways. A siPOOL targeting PLK-1 is therefore a valid control for transfection efficiency. The readout can be made by microscope or any standard assay to measure apoptosis.
You offer siPOOLs for human, mouse and rat. My target sequence is from a different species. Can I still order a siPOOL and how?
Yes you can!
All we need is some additional information:
the species you are working with
the host system you are working with
the target sequence
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