For 454 sequencing we usually use the Mint cDNA synthesis kit, Evrogen, that amplifies cDNA, with a modified OligodT (Chao Sun et al., 2010) and works pretty good

We have used the TargetAmp 2-round aRNA amplification kit 2.0 (Epicentre Biotechnologies) for two rounds of linear amplification of RNA from microdissected fungal samples. It is based on reverse transcription and in vitro transcription from the cDNA (modified Eberwine protocol) and should give a linear amplification (Teichert et al. 2012, BMC Genomics).

We use the Arcturus® RiboAmp® HS PLUS from life technologies. 2 rounds aRNA production

Hi. How much RNA do you need? The NUGEN Ovation PicoSL WTA V2 (for Affymetrix platform) is used for samples between 250 pg - 50 ng amount of total RNA and i can get around 6-8 ug cDNA(ss). Now i'm testing the Pico Profiling Kit from Sigma and from 25ng of total RNA, i can get 8-12ug cDNA(ds).

Your concern about getting a skewed data is very valid and this should be considered during analysis. However, in limiting situations amplifications is a necessity and not just an option. For that we had used TargetAmp 2-round aRNA amplification kit 2.0 (Epicentre Biotechnologies).They claim that amplification will have no effect on the relative expression values but that needs to be validated further. The same supplier have a 1 round amplification kit as well. The choice between the two is dependent on the starting material. Hopefully this chart will help

http://www.epibio.com/targetamp/targetamp_performance.asp#1

Hi there are lot o products available but depends what kind of platform you use. I have used Agilent labeling kit for Agilent arrays and it has a pretty good efficiency and amplification with 50ng it reaches to 5ug and is called the labelled cRNA.

I guess biases are there when you do do sequencing and micro arrays and do target amplification but still by visualization of data through different plots and normalizations help to remove the biases or technical error

Good luck

Hi we use Kreatech kit for the mRNA amplification. Hope that it will also work for you

Good Luck

I tried the Agilent too and its good if you have enough starting material (one round of amplification).

I have done a lot of work on this and we published it in our book

http://www.springer.com/biomed/molecular/book/978-1-61779-054-6

please see chapter 17

I tried many kits and ways and bias is always there. However, depend on the amount of tissue/mRNA you get your best bet is NuGen Whole transcriptome amplification system. Really works so well.

We have used MessageAmp II aRNA Amplification Kit (Life Technologies) that produced really good results. We used a whole genome microarray slide and a 2 channels method (agilent-like).

I had Nimblegen (RIP, btw) chips processed using the Sigma WTA2 TransPlex Complete Whole Transcriptome Amplification Kit with very good results. I would suggest that you check with your microarray supplier for validation data that they may have available; that was the case for Nimblegen and I suppose that other companies will have white papers and, if not, their tech support will have data on that.

We also used Message Amp II aRNA Amplification kit to work with Affymetrix microarrays. Every platform works differently, some of this kits might be mor suitable than others depending on what you are planning to do. RNA-Seq assays require a good quality RNA to start and a reasonable yield if you are thinking of depletion protocol prior library prep. Preamplification with RNA might be trick you then will need a good experimental design to make sure that in the analysis phase you have enough to correct the data from bias that preamp can introduce.

We would need that for Affy gene expression microarrays! Thanks all for your answers!

We are using the Applause WT-Amp St System + Encore Biotin Module from NuGEN for our Affy chips (Whole-Transcript-Expression)

I just posted an answer to a similar question on ResearchGate (https://www.researchgate.net/post/How_to_preamplify_mRNA_for_both_microarray_and_qRT-PCR). Here it goes:

I never got around to use the WT-Ovation kit from NuGEN but did read about the good success possible with this kit. A paper with a very systematic analysis of its performance can be found on BMC Research Notes, Vermeulen et al 2009 : https://www.biomedcentral.com/1756-0500/2/235

Best,

~a.