Recenly I use T3 megascript do in vitro transcription. The results is always bad, the OD260/280 is about 1.53. A big white pellet can be seen after isopropanol precipitation and RNA concentratin measure is about 1.0 ug/ul. However, when run a RNA gel, the band is very faint even using 5 uL. I don't know why? Maybe there is protein contamination? or RNAse digestion? Please help. Thanks a lot!
We normally do overnight reaction for in vitro transcription. Make sure your DNA template is good.
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【在 q**********0 的大作中提到】 : Recenly I use T3 megascript do in vitro transcription. The results is always : bad, the OD260/280 is about 1.53. A big white pellet can be seen after : isopropanol precipitation and RNA concentratin measure is about 1.0 ug/ul. : However, when run a RNA gel, the band is very faint even using 5 uL. I don't : know why? Maybe there is protein contamination? or RNAse digestion? Please : help. Thanks a lot!