Extract total RNA from as few as 1 cell
Karen Dolter Jeff Braman
The StrataPrep total RNA microprep kit provides a convenient method for isolating total RNA from small samples of cells. For the purification, RNA is bound to a solid support, which eliminates the need for organic extraction and alcohol precipitation. The resulting RNA is high quality, with no detectable DNA contamination and is suitable for subsequent quantitative or conventional RT-PCR.
Stratagenes recently released StrataPrep total RNA miniprep kit is designed to isolate total RNA from cells ranging from 105 to 107 cells. However, when you need total RNA from a smaller sample, the StrataPrep total RNA microprep kit is ideal. In the past, isolating total RNA from very small samples by traditional methods was difficult and inefficient because yields from small samples tend to be low due to losses during phenol-chloroform extraction and/or ethanol precipitation. Additionally, organic solvents are hazardous. Now, with Stratagenes StrataPrep total RNA microprep kit, total RNA can be extracted from as few as 10 cells to as many as 5 x 105 cells.
As shown in the Figure
1 overview, cultured cells are disrupted by guanidine thiocyanate,
then bound to a silica-based fiber matrix within a spin cup. RNA
is immobilized on the fiber matrix, which allows contaminants to be removed
while avoiding organic extractions and ethanol precipitation. DNA is removed
with a single DNase I incubation step directly on the matrix, the matrix
is washed, and pure RNA is eluted with a small volume of buffer. The recovered
RNA is then ready to use. The StrataPrep total RNA microprep kit provides