An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR.
Blackgram is an important pulse crop grown for its protein rich seeds. Its tissues are rich in polyphenols which hampered pure nucleic acid extraction and thus use of molecular techniques in breeding programme. In this study, a well established DNA extraction method for legumes, Gem-CTAB method has...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Indian Society of Plant Breeders
2019-03-01
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| Series: | Electronic Journal of Plant Breeding |
| Subjects: | |
| Online Access: | http://ejplantbreeding.org/index.php/EJPB/article/view/3122 |
| _version_ | 1819044981401190400 |
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| author | Avi Raizada and Souframanien Jegadeesan |
| author_facet | Avi Raizada and Souframanien Jegadeesan |
| author_sort | Avi Raizada and |
| collection | DOAJ |
| description | Blackgram is an important pulse crop grown for its protein rich seeds. Its tissues are rich in polyphenols which hampered pure nucleic acid extraction and thus use of molecular techniques in breeding programme. In this study, a well established DNA extraction method for legumes, Gem-CTAB method has been modified to isolate PCR compatible DNA from blackgram in a quick and economical way from a very small tissue. The amplification rate of DNA extracted through modified Gem-CTAB method was found to be high compared to Gem-CTAB method for begomovirus specific genes. While for RNA isolation, among different protocols, column based kit method followed by lithium chloride precipitation yielded high quality RNA for leaves compatible with RT-PCR. For flowers and roots, kit method alone or with its modifications yielded pure RNA. The above modified nucleic acid extraction method yielded PCR compatible DNA and RNA from phenolics rich blackgram tissues. |
| first_indexed | 2024-12-21T10:21:19Z |
| format | Article |
| id | doaj.art-7156114d96c9468bbaee95dc56f7bcde |
| institution | Directory Open Access Journal |
| issn | 0975-928X |
| language | English |
| last_indexed | 2024-12-21T10:21:19Z |
| publishDate | 2019-03-01 |
| publisher | Indian Society of Plant Breeders |
| record_format | Article |
| series | Electronic Journal of Plant Breeding |
| spelling | doaj.art-7156114d96c9468bbaee95dc56f7bcde2022-12-21T19:07:25ZengIndian Society of Plant BreedersElectronic Journal of Plant Breeding0975-928X2019-03-0110116717610.5958/0975-928X.2019.00020.6An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR.Avi Raizada andSouframanien JegadeesanBlackgram is an important pulse crop grown for its protein rich seeds. Its tissues are rich in polyphenols which hampered pure nucleic acid extraction and thus use of molecular techniques in breeding programme. In this study, a well established DNA extraction method for legumes, Gem-CTAB method has been modified to isolate PCR compatible DNA from blackgram in a quick and economical way from a very small tissue. The amplification rate of DNA extracted through modified Gem-CTAB method was found to be high compared to Gem-CTAB method for begomovirus specific genes. While for RNA isolation, among different protocols, column based kit method followed by lithium chloride precipitation yielded high quality RNA for leaves compatible with RT-PCR. For flowers and roots, kit method alone or with its modifications yielded pure RNA. The above modified nucleic acid extraction method yielded PCR compatible DNA and RNA from phenolics rich blackgram tissues.http://ejplantbreeding.org/index.php/EJPB/article/view/3122DNARNABlackgramModified Gem-CTABTRIzol reagent |
| spellingShingle | Avi Raizada and Souframanien Jegadeesan An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR. Electronic Journal of Plant Breeding DNA RNA Blackgram Modified Gem-CTAB TRIzol reagent |
| title | An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR. |
| title_full | An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR. |
| title_fullStr | An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR. |
| title_full_unstemmed | An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR. |
| title_short | An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR. |
| title_sort | improved method for rapid isolation of dna and rna from leaves flowers and roots of blackgram vigna mungo l hepper for detection of begomovirus infection and rt pcr |
| topic | DNA RNA Blackgram Modified Gem-CTAB TRIzol reagent |
| url | http://ejplantbreeding.org/index.php/EJPB/article/view/3122 |
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