| PETERKOVÁ, Martina, Irena KOUTNÁ, Lenka TESAŘOVÁ, Michaela POTĚŠILOVÁ, Michal KOZUBEK, Viera HRABČÁKOVÁ, Martin KLABUSAY, Michael DOUBEK and Jiří MAYER. Microarray analysis using limited amount of cells. Folia Biologica (Praha). 2009, vol. 2009, No 55, p. 53-60. ISSN 0015-5500. |
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@article{808351,
author = {Peterková, Martina and Koutná, Irena and Tesařová, Lenka and Potěšilová, Michaela and Kozubek, Michal and Hrabčáková, Viera and Klabusay, Martin and Doubek, Michael and Mayer, Jiří},
article_number = {55},
keywords = {cDNA microarray; RNA isolation; RNA amplifiacation; CD34+ cells},
language = {eng},
issn = {0015-5500},
journal = {Folia Biologica (Praha)},
title = {Microarray analysis using limited amount of cells},
volume = {2009},
year = {2009}
}
TY - JOUR
ID - 808351
AU - Peterková, Martina - Koutná, Irena - Tesařová, Lenka - Potěšilová, Michaela - Kozubek, Michal - Hrabčáková, Viera - Klabusay, Martin - Doubek, Michael - Mayer, Jiří
PY - 2009
TI - Microarray analysis using limited amount of cells
JF - Folia Biologica (Praha)
VL - 2009
IS - 55
SP - 53-60
EP - 53-60
SN - 00155500
KW - cDNA microarray
KW - RNA isolation
KW - RNA amplifiacation
KW - CD34+ cells
N2 - cDNA microarray technology is widely used in various biological and medical disciplines to determine gene expression profiles. Unfortunately, this technology requires a large quantity of input RNA. Since there is an increasing need to more precisely analyze defined cell sub-populations with low cell counts, working protocols using a minimal number of input cells are needed. Optimal RNA isolation and its accurate amplification are crucial to the success of these protocols. The HL60 cell line was used in the search for a suitable protocol that can be used for clinical samples of CD34+ hematopoietic cells obtained from bone marrow. The goal was to discover the best method of isolating and amplifying RNA from a small number of cells. Our evaluation of various methods and kits available in the market revealed that the combination of RNAqueous Kit (Ambion, USA) for RNA isolation and the SenseAmp Plus Kit (Genisphere, USA) for one-round RNA amplification produced the best results. This article presents a verified protocol describing a reliable and reproducible method for producing enough input RNA for microarray experiments when the number of cells is limited.
ER -
PETERKOVÁ, Martina, Irena KOUTNÁ, Lenka TESAŘOVÁ, Michaela POTĚŠILOVÁ, Michal KOZUBEK, Viera HRABČÁKOVÁ, Martin KLABUSAY, Michael DOUBEK and Jiří MAYER. Microarray analysis using limited amount of cells. \textit{Folia Biologica (Praha)}. 2009, vol.~2009, No~55, p.~53-60. ISSN~0015-5500.
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