hypervalent_iodine Posted January 12, 2014 Posted January 12, 2014 I don't know for sure, but I strongly suspect it's to make my life hell when I'm trying to do RNASeq work.
CharonY Posted January 14, 2014 Posted January 14, 2014 Well good thing that the manufacturers make a killing with kits for removal... Though at least for small genomes the throughput of even the MiSeq tends to be sufficient to push through the ribosomes (except maybe for low abundance RNAs, for which I distrust the quantitative data anyway...). With regards to OP, there are several potential reasons. One is the obvious fact that translation is the rate limiting step in protein production. If you had more mRNA but insufficient ribosomes they would not affect things much. 1
hypervalent_iodine Posted January 15, 2014 Posted January 15, 2014 Well good thing that the manufacturers make a killing with kits for removal... Though at least for small genomes the throughput of even the MiSeq tends to be sufficient to push through the ribosomes (except maybe for low abundance RNAs, for which I distrust the quantitative data anyway...). I continue to be absolutely amazed at how much biologists spend on those kits. The RiboMinus or Dynabeads kits I usually use for isolation tends to do the trick for most of my work, it's just that I work almost exclusively with totally random non-model species and they don't always like to do what the kits tell them to.
AndresKiani Posted January 15, 2014 Posted January 15, 2014 Cells have huge number of ribosomes, some have millions of Ribosomes, so that's why...
CharonY Posted January 15, 2014 Posted January 15, 2014 Cells have huge number of ribosomes, some have millions of Ribosomes, so that's why... that is a bit of a circular argument.... I continue to be absolutely amazed at how much biologists spend on those kits. The RiboMinus or Dynabeads kits I usually use for isolation tends to do the trick for most of my work, it's just that I work almost exclusively with totally random non-model species and they don't always like to do what the kits tell them to. Depletion is always an issue as it is not really as specific as one would hope them to be. I.e. sometimes you get different results if you deplete things with different kits, even if the target is supposedly the same. Worse for silencing assays, but I digress. And yes Mol Bio is a huge money devourer. Running such a lab is a constant rat race to get funds to spend on perishable enzymes, kits etc. If a student/postdoc screws up, it can put quite a dent in your budget. I do envy some less money intensive disciplines (e.g. bioinformatics).
Arete Posted January 15, 2014 Posted January 15, 2014 I do envy some less money intensive disciplines (e.g. bioinformatics). Ha - you (and my adviser) don't want to know what our monthly data storage bill just got to, but yes, it's nothing like the library prep cost for a 96 sample Illumina run. Data mining is much cheaper than generating novel data. The high performance computing department do want to know how I just crashed a 512GB node on the cluster. In their exact words "We're not upset, we just want to know what you were trying to do... [pairwise r2 calcs on a 6 x 105 SNP dataset from 108 individuals - forgot to auto delete non significant results from the out file]. So I guess we also break expensive toys, but usually not catastrophically.
CharonY Posted January 15, 2014 Posted January 15, 2014 Hah, reminds me of my first discussion about data storage for my instruments. The discussion was basically: "how much data do you generate?" "Let's see, I estimate about 4 gigs..." "Per month? that should be no issue whatsoever." "About every 3h with the mass spectrometry systems alone. I have not factored the sequencing facility in yet..." "K, we need a new budget." Yeah, they did not have omics people before...
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