4 replies to this topic

[cancerous]

Hi all,

Recently I isolated RNA from micro-dissected (LCM) mouse tissues using PicoPure RNA isolation kit. The problem though is that the amount of my RNA may not be enough for microarray analysis. I dissected only about 500-1500 cells and these are obviously not enuogh for microarray. I only have 10ul of RNA, so I do not want to waste it by measuring the concentration or checking its quality right now. However I'd just like to amplify it and then I may have to do some QC assys. Finally I want to submit it for microarray. Could anyone recommend me to get a good mRNA amplification kit for me? Thanks!

Edited by cancerous, 26 March 2009 - 07:46 AM.

[gogreen]

cancerous, on Mar 26 2009, 09:14 PM, said:

Hi all,

Recently I isolated RNA from micro-dissected (LCM) mouse tissues using PicoPure RNA isolation kit. The problem though is that the amount of my RNA may not be enough for microarray analysis. I dissected only about 500-1500 cells and these are obviously not enuogh for microarray. I only have 10ul of RNA, so I do not want to waste it by measuring the concentration or checking its quality right now. However I'd just like to amplify it and then I may have to do some QC assys. Finally I want to submit it for microarray. Could anyone recommend me to get a good mRNA amplification kit for me? Thanks!

Hi cancerous,
I'm not sure if any mRNA amplification kit would fit your need. Probably you should increase the amount of LCM tissue to start with. I've not done an LCM myself, But i've done RNA isolation from LCMs using Trizol and done microarray with that.

[mdfenko]

for gene expression microarrays, agilent has a "Low RNA Input Linear Amplification Kit"

[Geno]

cancerous, on Mar 26 2009, 10:14 PM, said:

Hi all,

Recently I isolated RNA from micro-dissected (LCM) mouse tissues using PicoPure RNA isolation kit. The problem though is that the amount of my RNA may not be enough for microarray analysis. I dissected only about 500-1500 cells and these are obviously not enuogh for microarray. I only have 10ul of RNA, so I do not want to waste it by measuring the concentration or checking its quality right now. However I'd just like to amplify it and then I may have to do some QC assys. Finally I want to submit it for microarray. Could anyone recommend me to get a good mRNA amplification kit for me? Thanks!

You can use Express art Amplfication kits for small amount of RNA

With the ExpressArt® mRNA Amplification Kits you can amplify nano micro and even picograms of RNA sufficient for multiple microarray experiments

For reference :

http://www.ocimumbio...ak_appnotes.asp

http://www.amp-tec.c...ublications.htm

Just to let you that we at GeneLogic- Ocimum Bisolution provides microarrays services with Agilent, Affymetrix and Illumina platform for expression studies, genotyping ,GWAS etc. We have just announce that we now have an enhanced range of services, new products and a brand new website (www.genelogic.com).

[Helios]

Nugen offers WT-ovation pico RNA amplification kits for amplifying as low as 500pg of rna which can be used for microarray.you can read the instructions on their site.
all the best for your experiment!!