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  2. → Viewing Profile: Topics: mahrak

mahrak

Member Since 05 Apr 2011
Offline Last Active May 21 2013 03:17 AM
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Topics I've Started

which to use: miR or miRexpression vector

21 May 2013 - 03:20 AM

hi everybody
I am going to transfect SW480 cell line with miR143 in order to prove the concept of "gain of function". and I was wondering if anybody could help me to make the dicission of using miR-143(as in oligonucleotide miR-143) or miRexpressin vectors.


any response it much appreciated in advance.

PS: developing a stable cell line is not an option

RNA journals

05 May 2013 - 06:46 AM

hello every body
I was wondering if any one could help me with a list of journals to which one can submitte an article on rna stability. I must say I just have a set of real time PCR data and GFP expression by flowcytomtry, obviously I am looking for a journal of low impact.



any reply is really appreciated

much smaller fragments after IVT

30 December 2012 - 10:52 PM

hey there all


I have been trying to do a round of In vitro transcription using TranscriptAid T7 High Yield Transcription Kit( fermentas):
1- I linearize my plasmid with BamHI and perform phenol/chloroform extraction followed by ethanol precipitation.
2- I set up the IVT reaction acording to manual and let sit at 37centigrade degrees for 2 hrs
3- I treat the reaction with DNase I ( kit supplemented)
4- I perform another round of phenol/chloroform extraction followed by ethanol precipitation to have Purified RNA
5- I check the concentration and purity by spect.
6- I run the produced RNA on gel along with an RNA ladder, unfortunately the size is much, much, much smaller than what I expect; desired RNA should be app. 1500base in lenght, what I observe on gel is 100base.
I already have checked for t7 termination sequences on my plasmid. there are none. so I expect the t7 polymerase to "run off".
HELP!!
HELP!!

in vivo naked-mRNA delivery

26 November 2012 - 01:55 AM

hi all
this might seem a rather-not-related subject as this is a miRNA forum, but I was wondering if any body has any kind of experince with in vivo naked-mRNA delivery, possibly using simple soltins like ringer's or PBS.

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