PCR-RFLP and Sequencing of TrnS/trnfM Fragment of Enhalus Acoriodes From Sanur Coastal Waters, Bali, Indonesia: a Preliminary Study

This study aimed to evaluate the resolution of PCR-RFLP and sequencing of trnS/trnfM in detecting genetic diversity of seagrass Enhalus acoroides at Sanur Beach, Sindhu Beach and Semawang Beach. This research used six samples E. acoroides from each location. DNA extraction method followed Doyle and Doyle. PCR amplifications were done using primers P672/P673, P674/P675 and ITS4/ITS5 followed by digestion using restriction enzymes EcoRI, PstI, HindIII, BamHI, Rsa, Mva and HinfI. Sequencing analysis used PCR products of primers trnS/trnfM. Testing of DNA sequences of E. acoroides were conducted using BLAST (Basic Local Alignment Search Tool). DNA sequences were further analyzed using MEGA 5.2 software (Molecular Evolutionary Genetic Analysis) to evaluate variations of DNA. The sequence alignments were done using ClustalW software to determine the homology between the DNA sequences. The results showed that 18 samples of E. acoroides from Sanur, Sindhu and Semawang Beach have no polymorphism based on restriction enzyme analyses. Furthermore, sequencing of trnS/trnfM region of 18 samples E. acoroides showed that the sequences were identical

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

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, SS

UA12/2/1 Hoops on the Hill

Special basketball edition of the College Heights Herald: Nations, Jeff. The Three Qs: Quickness, Quickness, Quickness Nations, Jeff. Sole Senior: Cypheus Bunton Leads by Example Varney, Dennis. Full-Court Pressure Varney, Dennis. After Sitting Out a Season, Gwen Doyle Ready for Action Topper Trivia Frakes, Jason. Jeff Rogers: Junior College All-American Feeling the Pressure Varney, Dennis. Foreign Player Adjusting to Western – Jaana Heikkila, International Students Western, University of New Orleans Top Sun Belt Polls 1993-1994 Basketball Rosters Lady Tops: Two Polls Place Team in Top Five Batters, Tom. Paul Sanderford Teaches More Than Basketball Nations, Jeff. Doubters Beware: Tops Will Be Better This Year Darnell Mee the Latest in a Long Line of Hilltopper Stars Varney, Dennis. Denise Hill, Gwen Doyle Should Guide Lady Tops to Tournamen

Optimization of DNA extraction of physic nut (Jatropha curcas) by selecting the appropriate leaf

Prayitno E, Nuryandani E. 2011. Optimization of DNA extraction of physic nut (Jatropha curcas) by selecting the appropriate leaf. Nusantara Bioscience 3: 1-6. Jatropha curcas L. has important roles as renewable source of bioenergy. The problem occurs on difficult of DNA extraction for its molecular breeding programs. The objectives of this research were to study which leaf best as source of DNA extraction. Four accession were used, namely J1 and J2 (Jawa Tengah), S1 (South Sumatra), and S2 (Bengkulu). First, third, fifth, seventh, and yellow leaves for each accession were extracted using modification of Doyle and Doyle (1987) method. Visualization and comparation with Lambda DNA, Spectrophotometer UV-Vis and cutting DNA with EcoRI enzyme were show quality and quantity of DNA. The result showed that third leaves have sufficient quality and quantity as source of DNA. Third leaves DNA quantity for J1 (19.33 µg/mL), J2 (26.21 µg/mL), S1 (31.20 µg/mL), dan S2 (61.03 µg/mL), and quality for each accession were 1.9063 (J1), 2.0162 (J2), 2.0116 (S1), and 2.0856 (S2)

The expanding toolkit of translating ribosome affinity purification

Translating ribosome affinity purification is a method initially developed for profiling mRNA from genetically defined cell types in complex tissues. It has been applied both to identify target molecules in cell types that are important for controlling a variety of behaviors in the brain, and to understand the molecular consequences on those cells due to experimental manipulations, ranging from drugs of abuse to disease-causing mutations. Since its inception, a variety of methodological advances are opening new avenues of investigation. These advances include a variety of new methods for targeting cells for translating ribosome affinity purification by features such as their projections or activity, additional tags and mouse reagents increasing the flexibility of the system, and new modifications of the method specifically focused on studying the regulation of translation. The latter includes methods to assess cell type-specific regulation of translation in specific subcellular compartments. Here, I provide a summary of these recent advances and resources, highlighting both new experimental opportunities and areas for future technical development.</jats:p

Report, July 25, 1954, Cleveland Police Department

Sergeant Lockwood and Detective Doyle reread all statements and reports regarding Marilyn\u27s murder and present 14 facts and corresponding contentions/analysis and a synopsis. They surmised that Sam\u27s affair with Susan Hayes could have been a motive for killing Marilyn, or at least a cause of the argument during which he lost his temper and killed her. They also proposed that Sam went down to the lake to wash off the blood on his clothing and staged a burglary after he had killed Marilyn

Report, July 25, 1954, Cleveland Police Department

Sergeant Lockwood and Detective Doyle reread all statements and reports regarding Marilyn\u27s murder and present 14 facts and corresponding contentions/analysis and a synopsis. They surmised that Sam\u27s affair with Susan Hayes could have been a motive for killing Marilyn, or at least a cause of the argument during which he lost his temper and killed her. They also proposed that Sam went down to the lake to wash off the blood on his clothing and staged a burglary after he had killed Marilyn