An improved method for the isolation of total RNA from Avicennia germinans leaves.
Isolation of high-quality RNA of Avicennia germinans L. tissue is difficult due to high levels of phenols and other substances that interfere when using conventional procedures for the isolation. These substances not only decrease the yield but also the quality of RNA is almost poor. We present here a simple RNA protocol and fast methodology that effectively removes these contaminating substances without affecting the yield. The protocol developed is based on the SDS/phenol method with modifications including beta-mercaptoethanol to prevent oxidation of phenolic complexes, and phenol/chloroform extraction is introduced to remove proteins, genomic DNA, and secondary metabolites, and co-precipitated polysaccharides. Both A260/A230 and A260/A280 absorbance ratios of isolated RNA were around 2 and the yield was about 0.3 mg g(-1) fresh weight. Good-quality total RNA from leaves of Avicennia germinans could be easily isolated within 2 h by this protocol which avoided the limitation of plant materials and could provide total RNA for all kinds of further molecular studies.
Gonzalez-Mendoza
Daniel
D
Departamento de Recursos del Mar, Cinvestav Unidad Merida, Merida, Yucatan, Mexico.
Moreno
Adriana Quiroz
AQ
Zapata-Perez
Omar
O
D032407:Avicennia / Q000235:genetics*; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D018515:Plant Leaves / Q000235:genetics*; D018749:RNA, Plant / Q000235:genetics* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D018744:DNA, Plant; D018749:RNA, Plant