scholarly journals Barcode DNA Edelweis (Anaphalis javanica) Berdasarkan Gen matK

Jurnal MIPA ◽  
2015 ◽  
Vol 4 (2) ◽  
pp. 131
Author(s):  
Muzakir Rahalus ◽  
Maureen Kumaunang ◽  
Audy Wuntu ◽  
Julius Pontoh
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Dna Barcode ◽  
Living Organism ◽  
Chain Reaction ◽  
Base Pairs ◽  
Barcode Dna ◽  
Reverse Primer ◽  
Total Dna ◽  
Polymerase Chain

DNA barcode merupakan metode identifikasi organisme hidup dengan menggunakan urutan DNA pendek (± 500 pasang basa). Tujuan dari penelitian ini adalah memperoleh barcode DNA Edelweis dan menganalisis kemiripan gen matK Edelweis (Anaphalis javanica) dengan kerabat terdekatnya. Isolasi DNA total Edelweis berhasil dilakukan dengan menggunakan  manual prosedur dari InnuPrep Plant DNA Kit yang dimodifikasi. Gen matK parsial telah diisolasi dengan metode Polymerase Chain Reaction (PCR) menggunakan Primer forward matK-1RKIM-f dan Primer Reverse matK-3FKIM-r. Hasil analisis sekuens menghasilkan barcode DNA edelweis berukuran 843 bp. Hasil analisis kemiripan menunjukkan tingkat kekerabatan terdekat dengan A. margaritaceae yaitu 99.86% pada BOLD System dan 100 % pada NCBI.DNA barcode is a method to identify living organism by using several short sequences of DNA (± 500 base pairs). The purpose of this study was to obtain a DNA barcode and analyze the similarity of matK genes of edelweis (Anaphalis javanica) with its closest relatives. Isolation of total DNA of edelweis has been succesfully done by using modified manual procedures of InnuPrep Plant Kit. matK partial gene has been isolated by the method of Polymerase Chain Reaction (PCR) using forward primer MATK-1RKIM-f and reverse primer MATK-3FKIM-r. Analysis of DNA sequences of edelweis confirmed its DNA barcode size was 843 bp. Furthermore, A. javanica showed similarity 99.86% in BOLD system and 100% in NCBI with A. margaritaceae.

scholarly journals Barcode DNA Tumbuhan Pangi (Pangium edule R.) Berdasarkan Gen matK

Jurnal MIPA ◽  
2014 ◽  
Vol 3 (2) ◽  
pp. 113
Author(s):  
Irmi Bangol ◽  
Lidya Irma Momuat ◽  
Maureen Kumaunang
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Dna Barcode ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Plant Dna ◽  
Barcode Dna ◽  
Total Dna ◽  
Polymerase Chain ◽  
Forward Primer

DNA barcoding merupakan suatu teknik yang digunakan untuk mempercepat dan mempermudah proses identifikasi organisme dengan menggunakan potongan gen tertentu. Penelitian ini bertujuan untuk menentukan sekuens DNA barcode tumbuhan pangi berdasarkan gen standar matK dan membandingkannya dengan spesies yang berkerabat dekat di GenBank. DNA total daun pangi diisolasi menggunakan Innuprep plant DNA kit dan berhasil diamplifikasi dengan proses Polymerase Chain Reaction (PCR) menggunakan primer berdasarkan gen matK. Hasil sekuensing fragmen DNA yang menunjukkan panjang 720 bp yang teramplifikasi oleh primer forward dan 780 bp untuk yang teramplifikasi oleh primer reverse. Hasil analisis BLASTn menunjukkan tingkat kemiripan tumbuhan pangi sangat tinggi dengan Trichadenia zeylanical, yaitu 99%, dan diikuti spesies lainnya (Kiggelaria africanal, 98%; Guthriea capensis, 96%; Acharia tragodes, 92%; Erythrospermum phytolaccoides, 92%; Hydnocarpus sp. Chase 1301, 90%; Carpotroche longifolia, 89%; Moultonianthus leembruggianus, 89% dan Pimelodendron zoanthogyne, 88%). Analisis komposisi asam amino menunjukkan bahwa matK Pangium edule dan kesembilan spesies tumbuhan lainnya bersifat hidrofobik.DNA barcoding is a technical used to accelerate and simplify the process identification of organism with by using a snipping of specific genes. This study aimed to determine the DNA sequences of plant barcoding standard pangi based gene matK and compare with closely related species in GenBank. Total DNA was isolated using Innuprep pangi leaf plant DNA kit and successfully amplified by the Polymerase Chain Reaction (PCR) using primers based on the gene matK. The results of sequencing long DNA fragments showed7 20 bp are amplified by the forward primer and 780 bp were amplified by the primer for reverse. Blast analysis of the results showed very extremely high the plant pangi degree of similarity with Trichadenia zeylanical, namely99%, and followed by other species (Kiggelaria africanal, 98%; Guthriea capensis, 96%; Acharia tragodes, 92%; Erythrospermum phytolaccoides, 92%; Hydnocarpus sp. Chase 1301, 90%; Carpotroche longifolia, 89%; Moultonianthus leembruggianus, 89% dan Pimelodendron zoanthogyne, 88%). Analysis of aminoacid composition showed that matK Pangium edule and nine other plant species are hydrophobic.

scholarly journals Isolasi dan Karakterisasi Barcode DNA Tanaman Nusa indah putih (Mussaenda frondosa) Berdasarkan Gen matK

Jurnal MIPA ◽  
2015 ◽  
Vol 4 (2) ◽  
pp. 111
Author(s):  
Jein Damanis ◽  
Lidya Irma Momuat ◽  
Meiske S. Sangi ◽  
Maureen Kumaunang
Keyword(s):  
Polymerase Chain Reaction ◽  
In Silico ◽  
Dna Barcode ◽  
In Silico Analysis ◽  
Chain Reaction ◽  
Plant Dna ◽  
Barcode Dna ◽  
Reverse Primer ◽  
Polymerase Chain ◽  
Silico Analysis

Telah dilakukan penelitian untuk menentukan urutan nukleotida dari barcode DNA tanaman Nusa indah putih berdasarkan gen matK dan mengkarakterisasi matK tanaman nusa indah putih dengan analisis in-silico. Isolasi DNA dari tanaman Nusa indah putih telah dilakukan berdasarkan manual prosedur dari InnuPrep Plant DNA kit yang dilanjutkan dengan amplifikasi dengan metode Polymerase Chain Reaction (PCR) menggunakan primer forward matK-1RKIM-f dan primer reverse matK-3FKIM-r kemudian dielektroforesis dan disekuensing. Sekuensing tanaman  Nusa  indah  putih  menghasilkan  kromatogram  yang  berkualitas  tinggi, dengan panjang sekuens 843 bp. Hasil Analisis in-silico menunjukan matK tanaman nusa indah putih bersifat basa, stabil, dan berinteraksi baik dengan air.A study to determine the nucleotide sequence of the DNA barcode Nusa indah putih plants based on gene matK and to characterize the matK of Nusa indah putih plant using in-silico analysis had been done. Isolation of DNA from Nusa indah putih plant was conducted by manual procedures of InnuPrep Plant DNA kit, followed by amplification using Polymerase Chain Reaction (PCR) method using matK-1RKIM-f as forward primer and matK-3FKIM-r as reverse primer then electrophoresis and sequenced. Nusa indah putih plants sequencing produced a high-quality chromatogram produce, with a length of 843 bp sequences. Results of in-silico analysis showed that matK Nusa indah putih plant is a base, stable, and well-interacted with water.

scholarly journals Barcode DNA berdasarkan Gen rbcL dan matK Anggrek Payus Limondok (Phaius tancarvilleae) (DNA Barcode of Payus Limondok Orchid (Phaius tancarvilleae) Based on the rbcL and matK genes)

2012 ◽  
Vol 4 (2) ◽  
Author(s):  
Beivy J Kolondam
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Dna Sequences ◽  
Dna Barcode ◽  
Universal Primers ◽  
Chain Reaction ◽  
Barcode Dna ◽  
Polymerase Chain ◽  
Matk Sequence ◽  
Close Relatives

Metode identifikasi spesies telah disepakati menggunakan barcode DNA standar yaitu gen rbcL dan gen matK. Tujuan penelitian ini untuk menentukan tingkat kemiripan sekuens barcode DNA tanaman Anggrek Payus Limondok (Phaius tancarvilleae) dengan spesies kerabatnya yang sudah terdata dalam BOLD Systems, merekomendasi penggunaan barcode untuk mengidentifikasi atau mengkonfirmasi spesies ini, dan mengamati variasi intraspesifik. Teknik Polymerase Chain Reaction (PCR) digunakan untuk mengamplifikasi sekuens gen rbcL dan matK melalui primer universal. Barcode rbcL menunjukkan kemiripan 100% (identik) dengan dua spesies berbeda dalam famili yang sama (Orchidaceae), sehingga tidak bisa diandalkan untuk identifikasi spesies P. tancarvilleae secara akurat. Sekuens matK sampel menghasilkan kemiripan 100% dengan spesies sama yang sebelumnya telah terdata dalam BOLD Systems. Kemiripan ini mengindikasikan rendahnya variasi genetik intraspesies tetapi sekuens matK dapat diandalkan untuk identifikasi atau konfirmasi spesies anggrek P. tancarvilleae. Kata Kunci: barcode, rbcL, matK, Phaius tancarvilleae Abstract Species identification methods convention have been recommended to use standard DNA barcode for plants; the rbcL and matK genes. The aims of this research were to determine similarities in barcode DNA sequences of Payus Limondok Orchid (Phaius tancarvilleae) with its close relatives that listed in BOLD Systems, to recommend the use of DNA barcodes for identification or confirmation of this species, and to observe intraspecific variations. Polymerase Chain Reaction technique was employed to amplify rbcL and matK genes using universal primers. The rbcL barcode of Payus Limondok resulted identical hit with other two different species in the same family (Orchidaceae), therefore, unreliable for accurate P. tancarvilleae species identification. The matK sequence of this plant was 100% similar with the same plant species listed in BOLD Systems. This similarity indicated low genetic variation within the species, but the matK sequence was found to be reliable for P. tancarvilleae orchid species identification or confirmation. Keywords: barcode, rbcL, matK, Phaius tancarvilleae

scholarly journals Penentuan Barcode DNA berdasarkan Gen matK dan Analisis In-silico MatK Rumput Macan (Lantana camara L.)

Jurnal MIPA ◽  
2015 ◽  
Vol 4 (1) ◽  
pp. 24
Author(s):  
Billy L. Mokoagow
Keyword(s):  
Polymerase Chain Reaction ◽  
In Silico ◽  
Dna Barcode ◽  
In Silico Analysis ◽  
Lantana Camara ◽  
Chain Reaction ◽  
Barcode Dna ◽  
Matk Gene ◽  
Polymerase Chain ◽  
Silico Analysis

DNA barcoding merupakan metode identifikasi spesies menggunakan potongan DNA pendek yang disebut barcode DNA. Gen matK merupakan gen standar untuk penentuan  barcode DNA tanaman. Penelitian ini bertujuan untuk menentukan barcode DNA tumbuhan rumput macan (L. camara L.) berdasarkan gen matK, serta melakukan analisis in-silico terhadap produk gen matK tumbuhan rumput macan (L. camara L.) dengan kerabat terdekatnya. Gen matK L. camara L. telah berhasil diamplifikasi dengan Polymerase Chain Reaction (PCR) menggunakan primer forward matK-1RKIM-f dan primer reverse matK-3FKIM-r. Analisis terhadap sekuens matK L. camara L. menunjukkan bahwa barcode DNA tumbuhan rumput macan (L. camara L.) terdiri dari 843 nukleotida. Selanjutnya, hasil analisis in-silico menunjukkan bahwa matK Lantana camara L. bersifat basa, stabil, dan dapat berinteraksi baik dengan air.DNA barcoding is a method of species identification using short pieces of DNA called DNA barcode. matK is a standard gene to determine DNA barcode of a plant. The aim of this research was to determine the DNA barcode of Rumput Macan plant (Lantana camara L.) based on matK gene, as well as in-silico analysis of the product matK gene Rumput Macan (L camara L.) with its closest relatives. L. camara L. matK gene was successfully amplified by Polymerase Chain Reaction (PCR) using forward primer MATK-1RKIM-f and reverse primer MATK-3FKIM-r. Analysis of the matK sequence of L. camara L. showed that the barcode DNA of rumput macan plant (L. camara L.) consisting of 843 nucleotides. Furthermore, the result of in-silico analysis showed that the matK of L camara L. is alkaline, stable, and able to interact well with water.

scholarly journals Identifikasi Barcode Tumbuhan Gedi Merah (Abelmoschus manihot L. medik) dan Gedi Hijau (Abelmoschus moschatus) Berdasarkan Gen matK

Jurnal MIPA ◽  
2014 ◽  
Vol 3 (2) ◽  
pp. 120
Author(s):  
Yusuf R. Fattah ◽  
Vanda S. Kamu ◽  
Max R. J. Runtuwene ◽  
Lidya I. Momuat
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Barcoding ◽  
In Silico ◽  
Dna Barcode ◽  
In Silico Analysis ◽  
Chain Reaction ◽  
Barcode Dna ◽  
Matk Gene ◽  
Polymerase Chain ◽  
Silico Analysis

Gedi (Abelmoschus L.) merupakan tumbuhan tropis. Tumbuhan ini memilki efek farmakologis. Masyarakat Minahasa mengkonsumsi daun gedi yang direbus tanpa diberi bumbu sebagai obat tradisional untuk menurunkan kadar kolesterol, antihipertensi dan antidiabetes. Suatu metode baru untuk mengidentifikasi dan menganalisis keanekaragaman genetika spesies telah dikembangkan dengan menggunakan potongan gen standar yang dikenal dengan teknik DNA barcoding. Salah satu gen yang terdapat pada tumbuhan yaitu gen matK telah digunakan sebagai gen standar untuk barcoding. Pada penelitian ini telah dilakukan isolasi DNA total dan gen matK penanda barcode DNA dari gedi merah dan gedi hijau, serta analisis in-silico terhadap produk gen matK gedi merah, gedi hijau, dan kerabat terdekatnya. Gen matK diisolasi dan diamplifikasi menggunakan metode Polymerase Chain Reaction (PCR) menggunakan primer forward (5’-CGTACAGTACTTTTGTGTTT ACGAG-3’) dan primer reverse (5’-ACCCAGTCCATCTGGAAATCTTGGTTC-3’). Hasil pengurutan nukleotida DNA barcode matK menunjukkan bahwa sebanyak 828 pb matK berhasil diisolasi untuk tumbuhan gedi merah dan tumbuhan gedi hijau. Urutan nukleotida matK gedi merah dan gedi hijau menunjukkan tingkat kemiripan yang tinggi, yaitu > 95%. Selain itu, hasil analisis in-silico menunjukkan bahwa protein MatK gedi dan kerabat terdekatnya bersifat hidrofobik.Gedi (Abelmoschus L.) is a tropical plant. This plant has the pharmacological effects. Minahasan people consumed boiled gedi without any spices addition to lower cholesterol level, blood pressure, and glucose level. A new method for identifying and analyzing the genetic diversity of species has been developed using standard gene known as DNA barcoding technique. One of the genes found in plants called matK gene was used as standard for DNA barcoding. In this research, identification of DNA barcode of red gedi and green gedi based on matK gene, and in-silico analysis on the matK gene products of red gedi, green gedi, and its closest relatives gedi have been done. matK gene was isolated with Polymerase Chain Reaction (PCR) using forward primer (5'-CGTACAGTACTTTTGTGTTTACGAG-3') and reverse primer (5'-ACCCAGTCCATCTGGAAATCTTGGTTC-3'). Barcode DNA of red and green gedi showed 828 bp nucleotide sequence based on matK gene. In addition, matK of both gedi showed high similarity, i.e. >95%. Furthermore, in-silico analysis of MatK gedi and its closest relative showed that this protein is hidrophobic.

scholarly journals Barcode DNA Anthurium Gelombang Cinta (Anthurium plowmanii) berdasarkan gen rbcL dan matK (The DNA Barcode of Anthurium Wave of Love (Anthurium plowmanii) based on rbcL and matK Genes)

2013 ◽  
Vol 3 (1) ◽  
Author(s):  
Beivy J Kolondam ◽  
Edy Lengkong ◽  
J Polii-Mandang ◽  
Runtunuwu Semuel ◽  
Arthur Pinaria
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Leaf Shape ◽  
Dna Barcode ◽  
Universal Primers ◽  
Rbcl Gene ◽  
Chain Reaction ◽  
Barcode Dna ◽  
Polymerase Chain ◽  
Close Relatives

AbstrakMetode standar untuk identifikasi spesies tumbuhan melalui barcode DNA telah direkomendasikan untuk menggunakan dua gen plastida yaitu rbcL dan matK. Tujuan penelitian ini yaitu untuk menentukan tingkat kemiripan sekuens barcode DNA tanaman Anthurium Gelombang Cinta (Anthurium plowmanii) dibandingkan spesies kerabatnya yang sudah terdata dalam BOLD Systems dan untuk merekomendasi penggunaan barcode DNA yang bisa diandalkan dalam mengidentifikasi spesies ini. Teknik Polymerase Chain Reaction (PCR) digunakan dalam perbanyakan sekuens fragmen gen rbcL dan gen matK oleh primer universal yang tersedia. Hasil penelitian menyimpulkan bahwa sampel tanaman A. plowmanii menghasilkan sekuens barcode rbcL yang mirip 100% (identik) dengan spesies A. cubense. Ini berarti barcode DNA rbcL tidak dapat digunakan untuk identifikasi tingkat spesies. Sekuens barcode matK sampel menunjukkan kemiripan 99,1% dengan A. ravenii yang berbeda dalam morfologi daun. Sekuens matK sampel bersifat unik diantara anggota-anggota genus Anthurium sehingga direkomendasikam penggunaannya untuk identifikasi sampai tingkat spesies.Kata Kunci: barcode, rbcL, matK, Anthurium plowmaniiAbstractStandard method for plant species identification through DNA barcode has been recommended to use two plastids genes; the rbcL and matK. The aims of this research were to determine similarities in DNA barcode sequences of Anthurium Wave of Love (Anthurium plowmanii) with its close relatives that listed in BOLD Systems and to recommend the reliable DNA barcode for identification of this species. Polymerase Chain Reaction was employed to amplify rbcL and matK genes fragments using available universal primers. The result showed that A. plowmanii sample was 100% similar to A. cubense. For that reason, the rbcL gene is not a reliable for species identification. Sequence of matK barcode showed 99.1% in similarity with A. ravenii which has different leaf shape. The matK sequence of sample was unique among all listed Anthurium members, therefore, this barcode are recommended for plant identification to the species level.Keywords: barcode, rbcL, matK, Anthurium plowmanii

Analysis of ancient bone DNA: techniques and applications

1991 ◽  
Vol 333 (1268) ◽  
pp. 399-407 ◽  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mitochondrial Dna ◽  
Dna Sequences ◽  
Genetic Information ◽  
Chain Reaction ◽  
Base Pairs ◽  
Dna Recovery ◽  
Human Femur ◽  
Ancient Bone ◽  
Polymerase Chain

The analysis of DNA from ancient bone has numerous applications in archaeology and molecular evolution. Significant amounts of genetic information can be recovered from ancient bone: mitochondrial DNA sequences of 800 base pairs have been amplified from a 750-year-old human femur by using the polymerase chain reaction. DNA recovery varies considerably between bone samples and is not dependent on the age of the specimen. We present the results of a study on a small number of bones from a mediaeval and a 17th-century cemetery in Abingdon showing the relation between gross preservation, microscopic preservation and DNA recovery.

Detection of the Glanzmann's Thrombasthenia Mutations in Arab and Iraqi-Jewish Patients by Polymerase Chain Reaction and Restriction Analysis of Blood or Urine Samples

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman
Keyword(s):  
Polymerase Chain Reaction ◽  
Base Pair ◽  
Restriction Analysis ◽  
Urine Samples ◽  
Chain Reaction ◽  
Base Pairs ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Base Pair Deletion ◽  
Polymerase Chain

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

scholarly journals Easy and Efficient DNA Extraction from Woody Plants for the Detection of Phytoplasmas by Polymerase Chain Reaction

Plant Disease ◽  
1999 ◽  
Vol 83 (5) ◽  
pp. 482-485 ◽  
Author(s):  
Margaret J. Green ◽  
Dan A. Thompson ◽  
Donald J. MacKenzie
Keyword(s):  
Polymerase Chain Reaction ◽  
Plant Material ◽  
Woody Plant ◽  
Leaf Roll ◽  
Chain Reaction ◽  
Efficient Procedure ◽  
Identical Manner ◽  
Total Dna ◽  
Polymerase Chain ◽  
Selection Of

A simple and efficient procedure for the extraction of high-quality DNA from phytoplasma-infected woody and herbaceous plants for polymerase chain reaction (PCR) detection is described. This procedure does not require phenol, chloroform, or alcohol for the precipitation of nucleic acids. Herbaceous and woody plant material are extracted in an identical manner with no additional purification or enrichment steps required. The method utilizes commercially available microspin-column matrices, and the extraction of total DNA can be achieved in less than 1 h. The method has been used to successfully purify phytoplasma DNA from whole leaves, leaf petioles and midribs, roots, and dormant wood from a diverse selection of plant material. The phytoplasmas detected by PCR include pear decline, western X-disease, peach yellow leaf roll, peach rosette, apple proliferation, Australian grapevine yellows, and Vaccinium witches'-broom.

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