mRNA-Seq protocol

[Xi Wang]

Hi all,

I am now working on RNA-seq data. I am wonderint whether the standard Illumina mRNA-Seq protocol is updated or not from Jan 2008 to Sep 2009. If updated, what's the difference?

Thanks in advance.

[antoniou]

The latest protocol on the Icon web site is dated from Sept 2009. I don't have the 2008 protocol so I cannot comment on the differences

[Xi Wang]

The latest protocol on the Icon web site is dated from Sept 2009. I don't have the 2008 protocol so I cannot comment on the differences

Thanks. I think the protocol before Sept 2009 was not mature, so it was not published. I am just wondering that the data generated by different labs would have different data quality, and some substaintial biases. I really appreciate it if you or others could give some comments on it.

[huguesparri]

2008 protocole and 2009 protocole are the same.
The only difference is that in the 2009 version, Illumina gives advices on how to extract/manipulate total RNA, asks for a RNA with RIN>8 and where in the protocole you can do Cold Storage option.

I joined the 2008 version to this message.

[Xi Wang]

2008 protocole and 2009 protocole are the same.
The only difference is that in the 2009 version, Illumina gives advices on how to extract/manipulate total RNA, asks for a RNA with RIN>8 and where in the protocole you can do Cold Storage option.

I joined the 2008 version to this message.

Thank you very much!

[Xi Wang]

Hi,

Anyone knows how to explain why the read distributions of a gene from two experiments are not the same (not the expression, even only one isoform)? It's normal or abnormal? Now we can discharge the protocol difference. I have no idea since I am not an experiment man. Thanks.