Optimization and comparison of different methods for RNA isolation for cDNA library construction from the reindeer lichen Cladonia rangiferina

Table 1 A comparison of different RNA isolation procedures

Method	Average Yield (μg/g of dry tissue)	Standard Deviation of Yield (μg/g)	A_260/280	A_260/230	Estimated extraction time for 1 mg of total RNA (hours)	Estimated cost for extracting 1 mg of total RNA (Euro)
Spectrum plant total RNA kit	31.2	22.4	2.01-2.05	2.00-2.27	54	979
TRIzol reagent	162.1	52.7	1.47-1.65	0.42-0.91	5	235
Dong & Dunstan	126.3	33.1	1.57-1.68	0.27-0.41	9	19
CTAB	69.3	19.4	1.40-1.56	0.58-1.01	9	27
CTAB + RNeasy	8.6	5.8	2.22-2.24	2.50-2.93	62	286

Shown is the method along with average nucleotide yield from starting material (μg/g) and the standard deviation from at least six extractions. The A_260/280 and A_260/230 ratios provide insight as to the quality of the resultant nucleotides - an extraction of sufficient quality for cDNA library construction should have ratio quality values of approximately 2.0. Also shown are estimates of time (h) and costs (Euro) required for the extraction of 1 mg of total RNA - this is a typically required amount for mRNA purification for cDNA library synthesis or for genome resequencing techniques.

ISSN: 1756-0500