hy, I am confused about the parameters which we consider during RNA concentration quantity. My first question is that what is range for high quality RNA ( 1.8-2.1 ul ) for q Rt-PCR and i got 1.9 concentration but graph results not so good. On the other hands my friend also get high quality RNA but his graph is better although concentration same like me. so any one please tell me that why its so different even value almost same. For further information i have attached my figures and also my friend so, it is easy to understand my question. Here all my result picture only Chinese writing form my friend picture.
You have consisantly high 230 absorbances which typically indicate extractant contamination (salts, phenol etc).
Though usually not a problem for qPCR, some downstream applications such as microarrays are sensitive to this kind of contamination as they can give higher background fluorescence.
http://www.bio.davidson.edu/gcat/protocols/NanoDrop_tip.pdf
It is hard to guess why there is a difference of yield, are you using same amount of tissue/cells, maybe he was more through with the breakdown of cells etc. You can do an extraction effiency calculation if you are worried you are not getting as much RNA as you think you can.
Dear Can firstly i want ti say you thanks and second thing is that i just want to know can we consider it as a good quality RNA for q RT PCR or not because as we know that the value between 1.8-2.1 is range for high quality so my RNA extracted conc. is 1.93. and you are right my friend extract RNA from Larvae and pupae of fruit flies while i was extracted from Antennae of female fruit flies after feed them Protein bait.
They should be fine for qPCR, i've had similar values for 260/230 which worked fine. From the attached graphs, your 260/280 from those graphs look okay as well. However sample 9 has noticable amount of protein contamination. Since you are likely to have replicates, I would just note this sample for downstream applications in case the result from this sample looks different to others.
Don't forget to check RNA integrity as well, then you can be really sure if it is good quality RNA.
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