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Improved Sensitivity for Staining RNA with SYBR Gold Stain

run on two identical gels and stained with either SYBR Gold or ethidium bromide. When total RNA was stained with SYBR Gold, the ribosomal RNA bands were clearly visible when as little as 10 ng of total RNA was run on the gel, and the Eagle Eye II image integrated at 5 frames at 1/30 of a second (Figure 2, Panel A). But, when total RNA was stained with ethidium bromide, a minimum of 50 to 125 ng of total RNA was required for detectin, even when the image was integrated at 30 frames at 1/30 of a second (Figure 2 Panel B).

Figure 2

Conclusions

Use the SYBR Gold stain combined with the Eagle Eye II imaging system to clearly visualize 250 ng of poly(A)+ RNA or less on a gel. When the amount of sample RNA is the limiting factor for making cDNA libraries, this combined procedure provides a vast improvement over the 1 g of RNA often required for ethidium bromide staining. Hence, it is now possible to run an aliquot of every RNA sample on a gel, even when the amount of available poly(A)+ RNA is limited, and still have enough RNA available for cDNA synthesis.

REFERENCES

  1. Chomczynski, P. and Sacchi, N. (1987) Anal. Biochem. 162: 156-159.

  2. Aviv, H. and Leder, P. (1972) Proc. Nat. Acad. Sci. USA 69: 1408-1412.


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