I tried them both and I would like to know which is better TRIzol vs Qiagen kit in terms of purification and high concentration.
As I have some primers that do not span more than one exon. I used classic TRIzol with the "phase lock gel tube" will this eliminate DNA contamination or would it be better to use Qiagen kit along with DNase kit?
Thank you
Hi Hala,
Typically, an expert in RNA extraction using TRIzol will produce better quantities of good-quality RNA than using a Qiagen RNA extraction kit. However, gaining this expertise is time-consuming as the learning curve is steep to get truly good at it. As the Qiagen RNA extraction kits very reliably produce good-quality RNA that is excellent for most downstream applications (sequencing, qPCR, etc), most people, including high-quality scientists, use the Qiagen extraction kits. The added cost of using Qiagen kits is negligible compared to the sheer time it takes to become truly good at TRIzol extraction.
Additionally, TRIzol is a phenol- and chloroform- based reagent and just plain dangerous to use. Avoiding the use of TRIzol is highly recommended if possible. If your adviser/boss needs convincing to buy the Qiagen kits, this is a very strong argument. If you do use TRIzol, make absolutely sure you take the safety warnings seriously - you really should be wearing the full lab coat, gloves, lab goggles, and work in a chemical fume hood.
If gDNA contamination is an issue, using the Qiagen gDNA elimination columns easily deals with it. Alternatively, some reverse transcription kits (such as the Qiagen RT2 kit) come with a gDNA digestion step that does the same thing prior to reverse transcription, and you wouldn't need to do it twice.
Hope this is useful!
Emily
Hi Hala,
Typically, an expert in RNA extraction using TRIzol will produce better quantities of good-quality RNA than using a Qiagen RNA extraction kit. However, gaining this expertise is time-consuming as the learning curve is steep to get truly good at it. As the Qiagen RNA extraction kits very reliably produce good-quality RNA that is excellent for most downstream applications (sequencing, qPCR, etc), most people, including high-quality scientists, use the Qiagen extraction kits. The added cost of using Qiagen kits is negligible compared to the sheer time it takes to become truly good at TRIzol extraction.
Additionally, TRIzol is a phenol- and chloroform- based reagent and just plain dangerous to use. Avoiding the use of TRIzol is highly recommended if possible. If your adviser/boss needs convincing to buy the Qiagen kits, this is a very strong argument. If you do use TRIzol, make absolutely sure you take the safety warnings seriously - you really should be wearing the full lab coat, gloves, lab goggles, and work in a chemical fume hood.
If gDNA contamination is an issue, using the Qiagen gDNA elimination columns easily deals with it. Alternatively, some reverse transcription kits (such as the Qiagen RT2 kit) come with a gDNA digestion step that does the same thing prior to reverse transcription, and you wouldn't need to do it twice.
Hope this is useful!
Emily
well, actually it depends on the which qiagen product that you use. Qigen has some kits that use organic extraction + filter system which means that you filter your trizol like extraction as a result, you take more pure RNA that includes even small RNAs.
But standart qiagen kit for RNA isolation, do not isolate
Enes is correct that the standard Qiagen extraction kit for RNA isolation does not capture the small RNAs. If you happen to be looking for small RNAs, using the Qiagen miRNA extraction kit would extract those.
Trizol get higher yield than column base kit but the purification is lower. Trizol get more protein and DNA contamination than RNA Easy Kit.
We have published a research regarding to Trizol, GENEzol and Column base kit.
Article Comparison of commercial RNA extraction kits and qPCR master...
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