Ovation RNA-seq system

basager · 03-18-2010, 07:28 AM

Have anyone tried the Ovation RNA-seq system from NuGen? I have small amounts of input RNA and consider to use this system to have enough starting material for RNA-seq. Any experience?

kainsteven · 06-10-2010, 09:32 AM

We have several conference posters and webinars posted on our web site to illustrate studies with low inputs of total RNA using the Ovation RNA-Seq System: http://www.nugeninc.com/nugen/index....na-seq-system/

Best,
Steve

ShannonJ · 06-23-2010, 10:50 AM

I love the NuGen kits. We routinely use them to generate random cDNA from viral RNA samples. If you follow with the Exon Module the products can be used for beautiful Illumina libraries.

kainsteven · 06-25-2010, 09:57 AM

It is no longer necessary to use the exon module to produce the double-stranded cDNA. We have packaged the complete solution for producing a dscDNA starting with total RNA in either an RNA-Seq or 3'-DGE experiment. Each of these workflows integrate directly with our Encore NGS Library Systems for the Illumina platform. Workflow time from total RNA to cBot is about 9 hours.

Steve

paul z · 07-27-2010, 08:38 AM

I'm having a difficult time understanding how, in the absence of a polyA selection or a ribosomal reduction method, and through the use of a mix of oligo dT and random primers, NuGEN is able to achieve such purportedly low % mapping to rRNA (~ 3-3.5%) -- are the random primers not_so_random, i.e., do they select against rRNA sequences?

Anyone try NuGEN in house and obtain similar metrics as reported in NuGEN's product literature? (http://www.nugeninc.com/tasks/sites/...ov_rna_seq.pdf)

Curious,
-Paul

ShannonJ · 07-27-2010, 01:21 PM

We've used them to sequence viruses along with host "background" sequence. With one channel of SE 76bp Illumina we get about 90% coverage of the viral genome, and little host rRNA.

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03-18-2010, 07:28 AM	#1
basager Junior Member Location: Oslo, Norway Join Date: Feb 2010 Posts: 3	Ovation RNA-seq system Have anyone tried the Ovation RNA-seq system from NuGen? I have small amounts of input RNA and consider to use this system to have enough starting material for RNA-seq. Any experience? Last edited by basager; 03-18-2010 at 08:39 AM.

06-10-2010, 09:32 AM	#2
kainsteven Member Location: SF Bay Area Join Date: Jul 2009 Posts: 16	Users of the Ovation RNA-Seq System We have several conference posters and webinars posted on our web site to illustrate studies with low inputs of total RNA using the Ovation RNA-Seq System: http://www.nugeninc.com/nugen/index....na-seq-system/ Best, Steve Last edited by kainsteven; 06-11-2010 at 12:57 PM.

06-23-2010, 10:50 AM	#3
ShannonJ Junior Member Location: Los Alamos, NM Join Date: Jun 2009 Posts: 3	I love the NuGen kits. We routinely use them to generate random cDNA from viral RNA samples. If you follow with the Exon Module the products can be used for beautiful Illumina libraries. __________________ /\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ Shannon L Johnson PhD Los Alamos National Laboratory Joint Genome Insitute shannonj@lanl.gov

06-25-2010, 09:57 AM	#4
kainsteven Member Location: SF Bay Area Join Date: Jul 2009 Posts: 16	Ovation RNA-Seq, 3'-DGE and Encore NGS Library Systems It is no longer necessary to use the exon module to produce the double-stranded cDNA. We have packaged the complete solution for producing a dscDNA starting with total RNA in either an RNA-Seq or 3'-DGE experiment. Each of these workflows integrate directly with our Encore NGS Library Systems for the Illumina platform. Workflow time from total RNA to cBot is about 9 hours. Steve

07-27-2010, 08:38 AM	#5
paul z Junior Member Location: New York, NY Join Date: Aug 2008 Posts: 7	I'm having a difficult time understanding how, in the absence of a polyA selection or a ribosomal reduction method, and through the use of a mix of oligo dT and random primers, NuGEN is able to achieve such purportedly low % mapping to rRNA (~ 3-3.5%) -- are the random primers not_so_random, i.e., do they select against rRNA sequences? Anyone try NuGEN in house and obtain similar metrics as reported in NuGEN's product literature? (http://www.nugeninc.com/tasks/sites/...ov_rna_seq.pdf) Curious, -Paul

07-27-2010, 01:21 PM	#6
ShannonJ Junior Member Location: Los Alamos, NM Join Date: Jun 2009 Posts: 3	We've used them to sequence viruses along with host "background" sequence. With one channel of SE 76bp Illumina we get about 90% coverage of the viral genome, and little host rRNA. __________________ /\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ Shannon L Johnson PhD Los Alamos National Laboratory Joint Genome Insitute shannonj@lanl.gov