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Questions related from Sebastian Vencken
I'm having some trouble obtaining an RNA 200-500 base fragment size in lysates from cells fixed with 1% formaldehyde. I've based the parameteres of my protocol on the Covaris protocol for Total...
11 November 2015 8,366 1 View
I was hoping to prepare a long term stock of PLGA in chloroform, which I can store at -20C. Would anybody know what the long term stability of PLGA in chloroform would be? Thanks
07 July 2015 6,879 4 View
I'm producing ~200 nm siRNA PLGA/DOTAP nanoparticles using the double-emulsion solvent evaporation technique. When I sized the particles using DLS the majority (95%) were of the correct size...
06 June 2015 7,228 9 View
The authors of the following paper make some excellent points about the use of and difference between repeats and replicates: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3321166/ They are very...
04 April 2014 2,387 13 View
I'm aiming to attempt the opposite of what many on this forum are trying. I'm seeking to perform Trizol extraction with the implicit aim to purify RNA species that are longer than small RNA...
02 February 2014 9,239 3 View
Would anybody perhaps know how an antibody compares to a short (~25 nt) oligonucleotide in terms of binding strength. I'm aware that this depends on many variables, but I'm just trying to get a...
07 July 2013 7,021 8 View
Seeking information on Reversible RNA-RNA crosslinking
07 July 2013 7,131 2 View
I intend to cross-link RNA using psoralens such as HMT. I would like to reverse these cross links, which I believe can be done with 254 nm UV or a base-catalysed reaction. I would like to create...
07 July 2013 7,978 0 View
I'm performing a RNA-precipitation protocol with specific biotin-TEG labelled probes which bind streptavidin magnetic beads. Over time my enrichments have become significantly lower. My suspicion...
06 June 2013 9,215 1 View
A number of groups use aptamer-conjugated RNA sequences to specifically isolate RNA binding proteins. What is the advantage of using aptamers over e.g. using oligonucleotides to pull-down those...
05 May 2013 6,179 5 View
Does anybody know the half-life of formaldehyde protein-protein, protein-RNA and protein-DNA crosslinks when the sample is incubated at 37C?
05 May 2013 8,555 11 View
I've come across various methods to determine the summarised error in qPCR experiments that included all biological and technical replicates. Although I haven't had much trouble performing these...
05 May 2013 4,929 27 View
I am trying to design 20 bp ssDNA oligos in order to strongly bind mRNAs with high specificity at room temperature or colder in high cation concentration (1 M). I'd like to avoid the formation of...
02 February 2013 4,006 11 View
I'm trying to validate a specific target of a specific miRNA by transfecting my cells by upregulating this miRNA with synthetic pre-miRs and using the 3'UTR sequence cloned with luciferase...
04 April 2012 584 9 View
Hi all, I would like to combine all my experimental and technical replicates into one value with a standard deviation. The easy part was calculating mean and standard deviation for each...
04 April 2012 5,490 6 View
Hi, I'm having trouble papers and reviews that show total miRNA expression to be lower in embryonic, adult and cancer stem cells compared to differentiated or normal cells. There's plenty out...
11 November 2011 6,700 0 View
11 November 2011 7,808 0 View
I'd really appreciate it. http://www.ncbi.nlm.nih.gov/pubmed/2438234 http://www.ncbi.nlm.nih.gov/pubmed/3204101 Many thanks
08 August 2011 4,960 1 View
Hi, I have two curves derived from a scatter plot. One is clearly linear and the other is non-linear. The data points on this scatter plot are uncertain (i.e. they have standard deviations). With...
08 August 2011 6,432 12 View
Hi, I've microarrayed samples after a gene knock-down (KD confirmed by qRT-PCR to be 95%) with Affymetrix Gene 1.0 ST arrays and Biotique XRAY software. The problem is, that this well expressed...
07 July 2011 5,852 0 View
