Genetic variability of Brazilian Toxoplasma gondii strains detected by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and simple sequence repeat anchored-PCR (SSR-PCR)

2004 ◽  
Vol 4 (2) ◽  
pp. 131-142 ◽  
Author(s):  
A FERREIRA ◽  
R VITOR ◽  
A CARNEIRO ◽  
G BRANDAO ◽  
M MELO
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Polymerase ◽  
Simple Sequence Repeat ◽  
Random Amplified Polymorphic Dna ◽  
Sequence Repeat ◽  
Chain Reaction ◽  
Rapd Pcr ◽  
Anchored Pcr ◽  
Polymerase Chain ◽  
Simple Sequence

DNA FINGERPRINTING OF SINGLE APHID EMBRYOS BY RANDOM AMPLIFIED POLYMORPHIC DNA POLYMERASE CHAIN REACTION (RAPD–PCR)

1999 ◽  
Vol 131 (2) ◽  
pp. 229-230 ◽  
Author(s):  
C.K. Chan ◽  
D.J. Petersen ◽  
T.C. Vrain
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Polymerase ◽  
Acyrthosiphon Pisum ◽  
Random Amplified Polymorphic Dna ◽  
Aphis Gossypii ◽  
Aphis Fabae ◽  
Chain Reaction ◽  
Rapd Pcr ◽  
Laboratory Colonies ◽  
Polymerase Chain

Extraction of DNA from whole aphids, in combination with random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) (Williams et al. 1990) markers can detect interspecific and intraspecific genetic variation (Black et al. 1992; Cenis et al. 1993). However, these techniques entail destructive sampling of fresh or preserved specimens. To allow experimental replication from a single sample while preserving the same aphid for morphometrical or karyotyping analyses, we describe a technique for RAPD-PCR using DNA from single aphid embryos. We evaluated the usefulness and reliability of single-embryo analysis, using four species of our laboratory colonies, namely Acyrthosiphon pisum (Harris), Aphis fabae Scopoli, Aphis frangulae group, and Aphis gossypii Glover.

scholarly journals METODE EKSTRAKSI DNA TANAMAN TANPA PRESIPITASI ETANOL UNTUK KEGIATAN POLYMERASE CHAIN REACTION (PCR)

2019 ◽  
Vol 6 (1) ◽  
pp. 29
Author(s):  
Kristianto Nugroho ◽  
Rerenstradika Tizar Terryana ◽  
. Reflinur ◽  
Puji Lestari
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Simple Sequence Repeat ◽  
Sequence Repeat ◽  
Chain Reaction ◽  
Ethanol Precipitation ◽  
Polymerase Chain ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing ◽  
Simple Sequence

A Simplified Plant DNA Extraction Protocol without Ethanol Precipitation for Polymerase Chain Reaction (PCR) Activities ABSTRACTMolecular-based research in agriculture includes DNA extraction stage involving DNA precipitation using ethanol or isopropanol which tends to take a long time. The purpose of this study was to obtain a plant DNA extraction method for Polymerase Chain Reaction (PCR) activities without going through the ethanol precipitation stage. Five important agricultural commodity crops, namely rice, corn, soybeans, chilies, and shallots were extracted by DNA using the modified Doyle and Doyle method. After the extraction phase using chloroform and isoamil alcohol solvents, the supernatant obtained was not precipitated using ethanol but was directly diluted and used as a template in PCR activities using two pairs of Simple Sequence Repeat (SSR) markers. The results showed that all samples could be well amplified, and amplicon tape visualized in both 1% agarose gel and 6% polyacrylamide gel were clearly visible. This method could save time and material, and reduce the dependence on liquid nitrogen. But this method is still limited to PCR requirements only, and cannot be used for activities that require high quality and quantity of DNA such as Next Generation Sequencing (NGS), digestion, and hybridization.Keywords: DNA extraction, ethanol precipitation, liquid nitrogen, PCR, SSR,  ABSTRAKPenelitian berbasis molekuler pada bidang pertanian mencakup tahapan ekstraksi DNA yang melibatkan presipitasi DNA menggunakan etanol atau isopropanol yang cenderung memakan waktu lama. Tujuan penelitian ini adalah untuk memperoleh metode ekstraksi DNA tanaman untuk kegiatan Polymerase Chain Reaction (PCR) tanpa melalui tahapan presipitasi etanol. Lima tanaman komoditas pertanian penting yaitu padi, jagung, kedelai, cabai, dan bawang merah diekstraksi DNA-nya menggunakan metode Doyle and Doyle yang dimodifikasi. Setelah tahap ekstraksi menggunakan pelarut kloroform dan isoamil alkohol, supernatan yang terbentuk tidak dipresipistasi menggunakan etanol melainkan langsung diencerkan dan digunakan sebagai template dalam kegiatan PCR menggunakan dua pasang marka Simple Sequence Repeat (SSR). Hasil menunjukkan bahwa seluruh sampel dapat teramplifikasi dengan baik serta pita hasil amplikon yang tervisualisasi baik pada gel agarosa 1% maupun gel poliakrilamid 6% terlihat jelas. Metode ini dapat menghemat waktu dan bahan serta mengurangi ketergantungan pemakaian nitrogen cair. Tetapi metode ini masih terbatas hanya untuk kebutuhan PCR saja dan tidak dapat digunakan untuk kegiatan yang membutuhkan DNA dengan kualitas serta kuantitas tinggi seperti Next Generation Sequencing (NGS), digesti, maupun hibridisasi.Kata Kunci: ekstraksi DNA, nitrogen cair, PCR, presipitasi etanol, SSR

scholarly journals Molecular typing of clinical and environmental isolates of Scedosporium prolificans by inter-simple-sequence-repeat polymerase chain reaction

2003 ◽  
Vol 41 (4) ◽  
pp. 293-300 ◽  
Author(s):  
M. Solé ◽  
J. Cano ◽  
J. L. Rodríguez-tudela ◽  
J. Pontón ◽  
D. A. Sutton ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Typing ◽  
Simple Sequence Repeat ◽  
Inter Simple Sequence Repeat ◽  
Sequence Repeat ◽  
Environmental Isolates ◽  
Chain Reaction ◽  
Scedosporium Prolificans ◽  
Polymerase Chain ◽  
Simple Sequence
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