Extraction of geminiviral DNA from a highly mucilaginous plant (Abelmoschus esculentus)

Joyce Jose, R. Usha

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

A protocol is described for the extraction of geminiviral DNA from bhendi yellow vein mosaic virus-infected Abelmoschus esculentus (known as bhendi or okra) containing high amounts of mucilage and other phenolic compounds. This method involves extraction with a buffer containing sodium citrate at pH 6 and PEG precipitation of the virus followed by alkali lysis. The extraction buffer eliminates the mucilage and other polyphenols, PEG precipitates the viral particles and DNA and the alkali lysis enriches the replicative forms of the viral DNA. The extracted DNA could be digested with restriction enzymes and cloned without any interference from chromosomal DNA. The quality of the DNA extracted by this method was compared to three other common plant DNA extraction protocols and was found superior. This method was used for PCR amplification and cloning of the 2.7 kbp DNA-A of BYVMV.

Original languageEnglish (US)
Pages (from-to)349-355
Number of pages7
JournalPlant Molecular Biology Reporter
Volume18
Issue number4
DOIs
StatePublished - Jan 1 2000

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Abelmoschus
Abelmoschus esculentus
DNA
Bhendi yellow vein mosaic virus
Viral DNA
Alkalies
Buffers
mucilages
Plant DNA
alkalis
Mosaic Viruses
buffers
Polyphenols
A-DNA
Virion
sodium citrate
Organism Cloning
Veins
okra
crossover interference

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Plant Science

Cite this

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abstract = "A protocol is described for the extraction of geminiviral DNA from bhendi yellow vein mosaic virus-infected Abelmoschus esculentus (known as bhendi or okra) containing high amounts of mucilage and other phenolic compounds. This method involves extraction with a buffer containing sodium citrate at pH 6 and PEG precipitation of the virus followed by alkali lysis. The extraction buffer eliminates the mucilage and other polyphenols, PEG precipitates the viral particles and DNA and the alkali lysis enriches the replicative forms of the viral DNA. The extracted DNA could be digested with restriction enzymes and cloned without any interference from chromosomal DNA. The quality of the DNA extracted by this method was compared to three other common plant DNA extraction protocols and was found superior. This method was used for PCR amplification and cloning of the 2.7 kbp DNA-A of BYVMV.",
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Extraction of geminiviral DNA from a highly mucilaginous plant (Abelmoschus esculentus). / Jose, Joyce; Usha, R.

In: Plant Molecular Biology Reporter, Vol. 18, No. 4, 01.01.2000, p. 349-355.

Research output: Contribution to journalArticle

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