High yield RNA extraction from Eucheuma denticulatum and Kappaphycus alveraazii (Gigartinales, Rhodophyta)
2006
Roohaida O. | Chan, P. C. | Diana M. N. | S., Lawrence E. K.
High quality RNA is a prerequisite in studying gene expression. However isolation of good quality RNA from algae is particularly difficult due to the high content of polysaccharides. To obtain large amounts of total RNA from Eucheuma denticulatum and Kappaphycus alvarezii, we compared the use of a commercial RNA extraction kit and a modified Lopez-Gomez and Gomez-Lim method. When applied to E. denticulatum, the modified Lopez-Gomez and Gomez-Lim method gave higher RNA yield of 57.4 µg per g of fresh tissue compared to 44.8 µg g-¹ obtaining using the commercial kit. Nevertheless, both methods produced RNA with high quality as determined by spectral analysis. Using the modified Lopez-Gomez and Gomez-Limmethod, much lower yield of RNA was obtained from K. avarezii albeit the same high quality. The quality of RNA extracted was also examined by reverse transcription-palymerase chain reaction (RT-PCR) and cDNA synthesis. These assays indicated that the RNA isolated is of high quality and suitable for further molecular analyses. Hence a cost saving protocol which gives high yield of total RNA with high quality has been developed.
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