CKC: isolation of nucleic acids from a diversity of plants using CTAB and silica columns

David C Henderson; John Hammond

doi:10.1007/s12033-012-9494-y

CKC: isolation of nucleic acids from a diversity of plants using CTAB and silica columns

Mol Biotechnol. 2013 Feb;53(2):109-17. doi: 10.1007/s12033-012-9494-y.

Authors

David C Henderson¹, John Hammond

Affiliation

¹ USDA-ARS, United States National Arboretum, Floral and Nursery Plants Research Unit, B-010A, Beltsville, MD 20705, USA. david.henderson@ars.usda.gov

PMID: 22271458
DOI: 10.1007/s12033-012-9494-y

Abstract

To assay for viruses in plant samples, we required a method for nucleic acid isolation that is rapid, simple, and applicable to the widest possible variety of plants. A protocol for isolation of total nucleic acid (TNA) was developed by combining common CTAB methods with silica spin columns. We report data on TNA purity and RNA quality from over 30 plant genera representing 25 families. Measurements showed that RNA is of high quality, and one-step RT-PCR was successfully performed on all samples. The protocol can be completed in less than 2 h.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Biotechnology / instrumentation
Biotechnology / methods*
Cetrimonium
Cetrimonium Compounds / chemistry*
Nucleic Acids / chemistry
Nucleic Acids / isolation & purification*
Plants / chemistry*
Plants / genetics
Potassium Acetate / chemistry
RNA, Plant / chemistry
RNA, Plant / isolation & purification
Reverse Transcriptase Polymerase Chain Reaction
Silicon Dioxide / chemistry*

Substances

Cetrimonium Compounds
Nucleic Acids
RNA, Plant
Silicon Dioxide
Potassium Acetate
Cetrimonium