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Optimization of a protocol
for extraction of Plasmodium falciparum RNA from
infected whole blood samples for use in DNA microarrays
Jaffu Chilongola1*, Sakurani Balthazary2
and Erasto Mbugi3
1Faculty
of Medicine, Kilimanjaro Christian Medical College, Tumaini
University, Tanzania.
2Department
of, Physiology, Biochemistry, Pharmacology and Toxicology,
Faculty of Veterinary Medicine, Sokoine University of
Agriculture, Tanzania.
3Department
of Biochemistry, School of Medicine, Muhimbili University of
Health and Allied Sciences (MUHAS), P.O. Box 65001, Dar es
Salaam, Tanzania.
*Corresponding author:
E-mail:
jchilx@yahoo.co.uk.
Tel : +255 762 162537.
Accepted 28 March, 2008 |
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This study was carried out
to determine the efficiency of two reagents, RNAlater and
RNAwiz, for their ability to stabilize Plasmodium
falciparum RNA in infected whole blood and saponin lysed
parasite pellets for use in DNA microarrays. Eight infected
blood samples were stored in each of the reagents, and RNA
extracted at days 0 and 56 post collection. RNA yields and
quality were compared at the different time points between
the two test reagents. We show that for both reagents,
higher RNA yields and quality is obtained when RNA is
isolated immediately after sample collection (day 0),
however, results show that RNAwiz
storage provides a marginally higher RNA yield compared to
RNAlater storage. Our results indicate that whole blood gave
slightly higher RNA yields with superior quality as compared
to saponin lysed samples when such whole blood samples are
stored in RNA wiz, but not in RNAlater. From our results, we
recommend RNAwiz as a better reagent for use in storage of
whole infected blood intended for extraction of P.
falciparum RNA for DNA microarrays and other sensitive
techniques.
Key words:
RNAwiz, RNAlater, RNA extraction, Plasmodium falciparum,
DNA microarrays. |
