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Fast Isolation of Total RNA from Small Numbers of Cells

ected from one tenth of the RNA sample isolated from a one-cell equivalent, with Ct values of 35 (HeLa) and 36 (THP-1) (Figure 6). As expected, the Ct value increases with a decrease in cell number.

Fig.6

Conclusions

The StrataPrep total RNA microprep kit is the method of choice for isolating RNA from small cell samples. The protocol and supplied reagents provide a method to purify pure, intact RNA with high yields. Amplification by conventional RT-PCR is feasible from as few as 1 cell and by quantitative RT-PCR from only one cell.

Acknowledgments

The authors thank Robert Saiz and Reinhold Mueller for performing molecular beacon RT-PCR and Sylvia Norman for assisting with conventional RT-PCR.

REFERENCES

  1. Sambrook, J., et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

  2. Steege, M. and Cayouette, M. (1996) Strategies 9: 53-56.

  3. Cayouette, M. and Hansen, C. (1996) Strategies 9: 56-57.

  4. Luckow, B., et al. (1994) Mol. Cell. Biol. 14: 1626-1634.

  5. Sabrouty, S. R.-E., et al. (1989) J. Mol. Evol. 29: 212-222.

  6. Piechaczyk, M., et al. (1984) Nature 312: 469-471.

  7. Borns, M., et al. (1999) Strategies 12: 33-36.

  8. Cayouette, M., et al. (1999) Strategies 12: 85-88.

  9. Cayouette, M., et al. (1999) Strategies 12: 89-92.

  10. Padmabandu, G. and Mueller, R. (1999) Strategies 12: 94-97.


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