Hi everyone,
I am a researcher at Brazil, working on translation regulation and I would like to know if someone have and could send me the rnai library clone efk-1?
The two commonly used RNAi libraries are the Ahringer RNAi library, and the ORFeome library, both distributed by Source BioScience. Most people will have obtained the libraries or clones from them under the terms of a Materials Transfer Agreement that frankly I don’t understand ("[the purchaser] shall not make the CLONES available to, or allow the use of the CLONES by (or for the benefit of), any person other than a Researcher carrying out investigation (as defined in the relevant MTA) and/or legitimate research" might be interpreted to allow distribution to anyone doing research?).
Fortunately, there’s a simple expedient: you can order a PCR primer pair for any large stretch of exonic sequence from your gene of interest, with T7 polymerase sites appended to the 5’ end of each primer. The PCR product can then be used to synthesize dsRNA for injection or soaking, or it can be cloned into any convenient vector (for example by T/A cloning) and transformed into the RNAi feeding strain HT115.
This doesn’t really scale well, but it’s cheaper and faster than ordering a single plasmid from the official sources, with no MTA issues.
We are also at the point where we are synthesizing stretches of DNA that we can’t get easily otherwise. gBlocks at IDTDNA.com have worked well for us. But for RNAi I agree with Hillel. In the past I have just amplified a stretch of genomic DNA with more than 100 bp of coding (exonic) sequence and dropped it into the opened L4440 plasmid.
You can also purchase them on a single clone basis from GE Dharmacon (Ugh, my stomach turns typing that oxybleep). For example:
http://dharmacon.gelifesciences.com/search/?term=R07E5.8
where R07E5.8 is the name for cku-80. RNAi of cku-80 is supposed to improve CRISPR knockin efficiency by compromising NHEJ. Here you see both feeding RNAi and ORFeome cDNA clones available. You can choose if it is worth the cost.
Edit: LOL @ “oxybleep”
Thanks a lot! I forgot the MTA