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Fast Isolation of Total RNA from Small Numbers of Cells


Extract total RNA from as few as 1 cell

Karen Dolter Jeff Braman
Stratagene

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.

Easy Protocol

Fig.1

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 all nece
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