GENETIC DIVERSITY EEL Anguilla marmorata BY RAPD IN THUA THIEN HUE PROVINCE, VIETNAM

Anguilla marmorata is a species with high economic value and increasingly interested by organizations and scientists. So far, many of the eel's biological characteristics remain mysterious, and they are often classified according to morphological features such as pigmentation patches, number of vertebrae, ... It is even difficult to distinguish one individual from another in some species, especially in the larval stage. In this study, the Random amplification of polymorphic DNA (RAPD) molecular marker was used to evaluate the genetic diversity of 48 eels collected in Thua Thien Hue province. Results showed that the genetic diversity of individuals in the Eel population studied is quite high. With 8 random primers via PCR, 77 DNA tapes with 76 polymorphic tapes were obtained, the tape size ranged from 170-2,500 bp, in which primer S10 showed the highest diversity with an average Ho value of 0.563, followed by primer S8 (Ho = 0.558). The lowest diversity was in the OPD5 primer (Ho = 0.300). The OPG17 primer is the primer that produces the most polymorphic tapes (13/13 tapes) and the S3 primer for the least amplified tapes polymorphism (9/10 DNA tapes). The diversity coefficient in each random primer ranged from about 0.300 to 0.563, with an average of 0.433. The genetic variation in the Eel population is random. Genetic variation can be attributed mainly to different eel breeding conditions and origins. Genetic similarity coefficients among the Eels varied from 0.660 to 0.910 and were divided into two main groups in genetic similarity coefficient 0.660.

KERAGAMAN GENETIK KELAPA DALAM MAPANGET NOMOR 32 HASIL PENYERBUKAN SENDIRI BERDASARKAN PENANDA RAPD

<p><strong>Genetic variability of selfing Mapanget Tall coconut No. 32 based on RAPD marker</strong></p><p>The objective of this study was to analyze the genetic variability of seling Mapanget Tall coconut No. 32 used RAPD marker. Ihe method of morphology, fruit component and isozymc analyses could not explain the homozygote level of the offspring and his parents. The DNA analysis was done at Plant Biology Laboratory, PAU, Life Science IPB, and the coconut leaflets samples were collected from Kima Atas Research Instalation, Research Institute for Coconut and Palmae, Manado. This research the study was conducted from November 1999 to Fcbruay 2000. Coconut materials analyzed were DMT 32-OP, DMT 32-S2, DM 1 32-S3 andDMT32-S4. DMT 32-S2 was the offsprings of the selfcd DMT 32- SI coconut seling. and the DMT 32-SI came from the sclfed of DMT 32-OP. Then selfcd DMT 32-S2 produced were found the DMT 32-S3. and selfcd of DMT 32-S3 produced the progeny of DMT 32-S4. DNA was isolated using the method of ROHDE et at (1995), while DNA quantity and quality was using the method of SAMBROOK et al (1989). Then the DNA was ampliied using 10 random primer 10 mer and PCR apparatus of 2.400 Perkin-Elmer System. Ater thai the DNA was elcctrophorated, and photographed using Polaroid 667 ilm, then ihe biner data matrix of each coconut population was calculated lor Ilic number of monomorphism banding was found in DMT 32-S2. Ihe genetic similarity between DMT 32-S3 and DMT 32-S3 was the mosl similar at genetic distance of 90%. DMT 32-S3 resulted from self pollination can be recommended as parent material for hybridization.</p>

Kemiripan genetik wereng coklat, Nilaparvata lugens Stål. (Homoptera: Delphacidae) populasi Klaten dan Yogyakarta berdasarkan penanda RAPD-PCR

Wereng coklat (Nilaparvata lugens Stål.) populasi Klaten dan Yogyakarta menunjukkan kemampuan adaptasi pada varietas padi tahan lebih cepat daripada populasi asal wilayah sekitarnya, namun kajian aspek genetik terkait hal ini masih terbatas. Penelitian ini dilakukan untuk mengidentifikasi kemiripan genetik wereng coklat populasi Klaten dan Yogyakarta dengan populasi asal lokasi sekitarnya sebagai pembanding, berdasarkan penanda RAPD-PCR. Pengambilan sampel wereng coklat dilakukan di pertanaman padi di Klaten, Yogyakarta, Sukoharjo, Boyolali, Karanganyar, Sragen, dan Ngawi. Identifikasi kemiripan genetik dilakukan dengan teknik RAPD-PCR, menggunakan lima random primer, yakni OPB 01, OPB 07, OPC 04, OPC 08, dan OPN 15. Hasil pengujian menunjukkan bahwa kelima primer mampu mengamplifikasi DNA wereng coklat dengan baik, namun tidak ada primer yang mampu membedakan secara jelas populasi Klaten dan Yogyakarta dengan populasi asal wilayah sekitarnya. Populasi wereng coklat asal Klaten dan Yogyakarta juga menunjukkan kemiripan genetik dengan populasi asal wilayah sekitarnya, yakni Boyolali, Sukoharjo, dan Sragen, kecuali dengan Karanganyar dan Ngawi. Kajian genetik antar populasi, termasuk populasi asal Klaten dan Yogyakarta diperlukan untuk mengungkap perbedaan genetiknya.

A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism

The clustered regularly interspersed palindromic repeats (CRISPR)/Cas9 (CRISPR-associated protein 9) system is a powerful tool for genome editing that can be used to construct a guide RNA (gRNA) library for genetic screening. For gRNA design, one must know the sequence of the 20-mer flanking the protospacer adjacent motif (PAM), which seriously impedes experimentally making gRNA. I describe a method to construct a gRNA library via molecular biology techniques without relying on bioinformatics. Briefly, one synthesizes complementary DNA from the mRNA sequence using a semi-random primer containing a PAM complementary sequence and then cuts out the 20-mer adjacent to the PAM using type IIS and type III restriction enzymes to create a gRNA library. The described approach does not require prior knowledge about the target DNA sequences, making it applicable to any species.