The Chemistry Behind Plant DNA Isolation Protocols

AbstrakProtokol dan metode sederhana, efisien untuk isolasi DNA genom tanaman durian yang banyak mengandung phenol dan residu polisakarida telah dihasilkan. Pada penelitian ini, digunakan protokol isolasi DNA tumbuhan dengan metode CTAB yang dimodifikasi sebagai protokol yang efisien untuk membuang polisakarida, phenol dan lendir yang sangat melimpah pada tanaman durian. Obyek penelitian ini terdiri dari protocol CTAB yang dimodifikasi tahap inkubasi dan presipitasi pemurnian DNA genom dari phenol dan polisakarida. Perbandingan 2 protokol isolasi DNA durian dengan CTAB standard an CTAB modifikasi menunjukkan bahwa metode CTAB modifikasi menghasilkan whole genom durian cukup murni rata rata 1,99 dan berhasil diamplifikasi dengan PCR-RAPD.Kata kunci: isolasi, DNA, polisakarida, CTAB, modifikasi, . AbstractThe simple and efficient method for genomic DNA isolation protochol from durian , its woody fruit crops containing high polysaccharide levels has been described here. In the present study, using modified CTAB for plant DNA isolation protocols were studied for removing the highly concentrated polysaccharides from genomic DNA of woody fruit crops.This method involves the modified CTAB at the incubate and precipitate procedure employing DNA purification step to remove polysaccharides and phenol residu. Compared with the two studied DNA isolation protocols of durian using standart CTAB and modified CTAB the everage yield high quality DNA whole genom is 1,99 purity and DNA was suitable for PCR and RAPD analyses.Keyword: isolation, DNA, polysaccharides, phenol residu, CTAB

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Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides, and RNA

Taxon ◽

10.2307/1223408 ◽

1995 ◽

Vol 44 (3) ◽

pp. 379-386 ◽

Cited By ~ 58

Author(s):

David V. Jobes ◽

David L. Hurley ◽

Leonard B. Thien

Keyword(s):

Dna Isolation ◽

Plant Dna

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Quick Miniprep for Plant DNA Isolation

Cold Spring Harbor Protocols ◽

10.1101/pdb.prot5179 ◽

2009 ◽

Vol 2009 (3) ◽

pp. pdb.prot5179-pdb.prot5179 ◽

Cited By ~ 8

Author(s):

D. Weigel ◽

J. Glazebrook

Keyword(s):

Dna Isolation ◽

Plant Dna

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Dellaporta Miniprep for Plant DNA Isolation

Cold Spring Harbor Protocols ◽

10.1101/pdb.prot5178 ◽

2009 ◽

Vol 2009 (3) ◽

pp. pdb.prot5178-pdb.prot5178 ◽

Cited By ~ 1

Author(s):

D. Weigel ◽

J. Glazebrook

Keyword(s):

Dna Isolation ◽

Plant Dna

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Plant DNA Isolation Protocols

DNA Fingerprinting in Plants ◽

10.1201/9781420040043.ax1 ◽

2005 ◽

pp. 311-321

Keyword(s):

Dna Isolation ◽

Plant Dna

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A Plant DNA Isolation Protocol Suitable for Polymerase Chain Reaction Based Marker‐Assisted Breeding

Crop Science ◽

10.2135/cropsci1998.0011183x003800010036x ◽

1998 ◽

Vol 38 (1) ◽

pp. 217-220 ◽

Cited By ~ 12

Author(s):

D. A. Lange ◽

S. Peñuela ◽

R. L. Denny ◽

J. Mudge ◽

V. C. Concibido ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Dna Isolation ◽

Marker Assisted Breeding ◽

Chain Reaction ◽

Plant Dna ◽

Polymerase Chain ◽

Dna Isolation Protocol

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DNA Isolation And Amplification of the rbcL (ribulose-1,5- bisphosphate carboxylase/oxygenase large subunit) gene of Caulerpa sp., Gracilaria sp., And Sargassum sp.

JURNAL ILMIAH PLATAX ◽

10.35800/jip.8.2.2020.30003 ◽

2020 ◽

Vol 8 (2) ◽

pp. 214

Author(s):

Biondi Tampanguma ◽

Grevo S. Gerung ◽

Veibe Warouw ◽

Billy Th Wagey ◽

Stenly Wulllur ◽

...

Keyword(s):

Target Gene ◽

Dna Isolation ◽

Large Subunit ◽

Dna Amplification ◽

Ammonium Bromide ◽

Rbcl Gene ◽

Bisphosphate Carboxylase ◽

Plant Dna ◽

Ctab Method ◽

Amplification Process

DNA isolation and gene amplification of algae are significantly influenced by various factors such as characteristics and components of the algae cell wall. Therefore techniques and methods of DNA isolation in certain algae, sometimes only succeed in one particular species and can not be applied to another algae species. Based on that issue, this study was conducted with the aims to determine the succeed of DNA isolation and amplify the rbcL gene as a target gene for identification. Algae DNA was isolated by using innuPrep Plant DNA commercial kit, and the second one with a modified conventional Cetyl Trimetyl Ammonium Bromide (CTAB) method, for the amplification process was using rbcL gene (ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit) with two pairs of primers : rbcL 7F-753R and rbcL 577F-rbcSR. The results showed that the DNA of Gracilaria sp was succeed isolated by using CTAB method and it was denoted by the presence of DNA bands in agarose gel. Meanwhile DNA amplification for Gracilaria sp., and Sargassum sp., were succeed amplified with the appearance of DNA bands. But in algae Caulerpa sp., was only succeed on 1 pair of primary rbcL 7F and 7.Keywords : DNA, gene rbcL, algae Caulerpa sp., Sargassum sp., Gracilaria sp;AbstrakIsolasi DNA dan amplifikasi gen pada alga sangat dipengaruhi oleh beberapa faktor seperti karakter dan komponen pada dinding sel alga. Oleh karena itu proses isolasi DNA terkadang bisa berhasil pada satu jenis alga, namun tidak berhasil pada jenis alga lainnya. Oleh karena alasan tersebut, maka penelitian ini dilakukan untuk menentukan keberhasilan Isolasi DNA dan mengamplifikasi gen rbcL sebagai gen target identifikasi. Penelitian ini dilakukan dengan tahapan awal Isolasi DNA yang menggunakan kit komersil innuPrep Plant DNA Kit, dan metode konvensional Cetyl Trimetyl Ammonium Bromide (CTAB) yang telah dimodifikasi. Sedangkan untuk proses amplifikasi, menggunakan gen rbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit) digunakan dua pasang primer yaitu rbcL 7F-753R dan rbcL 577F-rbcSR. Hasil isolasi DNA dari alga Gracilaria sp berhasil diisolasi menggunakan metode CTAB yang ditandai dengan adanya pita DNA pada gel agarose. Amplifikasi DNA pada alga Gracilaria sp., dan Sargassum sp., berhasil diamplifikasi dengan munculnya pita DNA. Namun pada alga Caulerpa sp. hanya berhasil pada 1 pasang primer rbcL 7F dan753R.Kata kunci : DNA, gen rbcL, alga Caulerpa sp., Sargassum sp., Gracilaria sp.

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Genomic DNA isolation from scrophularieae dried leaves using a simple, high-throughput protocol

Bangladesh Journal of Botany ◽

10.3329/bjb.v48i4.49083 ◽

2019 ◽

Vol 48 (4) ◽

pp. 1231-1235

Author(s):

Mehrshid Riahi ◽

Melina Babaei ◽

Farrokh Ghahremaninejad

Keyword(s):

Molecular Weight ◽

High Molecular Weight ◽

Dna Extraction ◽

High Throughput ◽

Genomic Dna ◽

Dna Isolation ◽

High Molecular Weight Dna ◽

Plant Dna ◽

Genomic Dna Isolation

This communication described efficient DNA extraction from Scrophularia and Verbascum samples. Modified Murray and Thompson modified Cota-Sànchez method and Bioflux kit methods were applied for the extraction of DNA. Among the different methods, Bioflux kit Plant DNA extraction kit, coupled with some modification was the best for extraction of high molecular weight DNA as long as the extracted DNA is quantified with fluorescence-based methods.

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