I think there is some confusion, here. Hoechst dyes bind to the minor groove of double stranded DNA, mostly in AT rich sequences. The binding to RNA is highly inefficient, and would occur only in the presence of secondary structure of the RNA.

To quantify RNA a a UV spectrophotometer is sufficient. For quantification the reading should be done at 260 nm wavelength. What is important for quantification is that at this wavelength 40 microg of RNA in 1 ml volume in a quartz cuvette will give you absorbance of 1 with a light path of 1 cm. Apart from the 260-230 nm ratio, which is an indication of chemical purity of your RNA from sugars and/or guanidinium (in case you used Trizol to extract), the 260/280 nm ratio is also very important to determine chemical purity from phenol (and partially from proteins).

The Nanodrop is a good idea, if you have access to it, but any spectrophotometer will do, you might only need to use up more RNA due to minor sensitivity compared to a Nanodrop (there are also cuvettes for 100 microl volume reading instead of 1 ml).

A Qubit is indeed a nice instrument that will allow you to select for dsDNA, ssDNA or RNA by using different dyes... a lot less expensive than a Nanodrop, might be more accessible.

In any case, you should always load on gel 0.2-0.5 microg of RNA to verify its integrity. The two ribosomal RNA bands should be clearly visible, depending from the organism you are working with they will run a bit differently and their intensity ratio will also be slightly different, usually between 2 and 2.5 (bigger isoform vs smaller isoform). If this ratio is lower you have extracted degraded RNA and you should evaluate if it is worth using it...

Alternatively you can use a Bioanalyzer that will quantify and provide you an integrity ratio at the same time... But this is a very expensive instrument, so not easy to access...

if you have a nanodrop or qiaxpert then no need to put any dye, check 260.230 ratio it should be 2.0 to 2.2, it fou want more accurate reading go for qubit fluorescence 

I think there is some confusion, here. Hoechst dyes bind to the minor groove of double stranded DNA, mostly in AT rich sequences. The binding to RNA is highly inefficient, and would occur only in the presence of secondary structure of the RNA.

To quantify RNA a a UV spectrophotometer is sufficient. For quantification the reading should be done at 260 nm wavelength. What is important for quantification is that at this wavelength 40 microg of RNA in 1 ml volume in a quartz cuvette will give you absorbance of 1 with a light path of 1 cm. Apart from the 260-230 nm ratio, which is an indication of chemical purity of your RNA from sugars and/or guanidinium (in case you used Trizol to extract), the 260/280 nm ratio is also very important to determine chemical purity from phenol (and partially from proteins).

The Nanodrop is a good idea, if you have access to it, but any spectrophotometer will do, you might only need to use up more RNA due to minor sensitivity compared to a Nanodrop (there are also cuvettes for 100 microl volume reading instead of 1 ml).

A Qubit is indeed a nice instrument that will allow you to select for dsDNA, ssDNA or RNA by using different dyes... a lot less expensive than a Nanodrop, might be more accessible.

In any case, you should always load on gel 0.2-0.5 microg of RNA to verify its integrity. The two ribosomal RNA bands should be clearly visible, depending from the organism you are working with they will run a bit differently and their intensity ratio will also be slightly different, usually between 2 and 2.5 (bigger isoform vs smaller isoform). If this ratio is lower you have extracted degraded RNA and you should evaluate if it is worth using it...

Alternatively you can use a Bioanalyzer that will quantify and provide you an integrity ratio at the same time... But this is a very expensive instrument, so not easy to access...

Qubit works wonderfully, no doubt about that.

Hello friends,

Thank you for your responses. What about using Quant it RNA detection kit.

can anyone help me with this.

Thank you.