Biomedical Engineering Reference
In-Depth Information
RNA Amplification Strategies:
Toward Single-Cell Sensitivity
Natalie Stickle
1
, Norman N. Iscove
2
,CarlVirtanen
1
, Mary Barbara
2
,
Carolyn Modi
1
,ToniDiBerardino
3
, Ellen Greenblatt
3
,TedBrown
4
and
Neil Winegarden
1
1
University Health Network Microarray Centre, Toronto, Ontario, Canada
2
Ontario Cancer Institute, Princess Margaret Hospital, University Health Network, Toronto, Ontario,
Canada
3
Mount Sinai Hospital Centre for Fertility and Reproductive Health, Toronto, Ontario, Canada
4
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Joseph and Wolf Lebovic Centre, Toronto,
Ontario, Canada
5.1 Introduction
Microarray technology has largely become commonplace in genomics research initiatives,
and yet, despite the major potential of the technology, there has been debatable success
from the many microarray experiments that have thus far been performed. One of the
key contributors to the mixed review of microarray technology has been the fact that the
limitations of the technology have largely, to date, limited the types of experiment that can
be performed. In particular, owing to the fairly high sample requirements for microarray
technology, researchers have resorted to the study of bulk, heterogeneous tissues which
have obfuscated any meaningful signatures that might have been obtained if more pure
populations of cells had been used.
5.1.1 The need for amplification
The first microarray protocols published called for in excess of 10
μ
g of total RNA and in
many of the earliest publications, as much as 100-500
g of total RNA [1, 2]. Given that
a single mammalian cell might contain from 2 to 35 pg of total RNA (Table 5.1), these
μ
