Development of Pinus koraiensis SSR Primers Based on EST-SSR Information Technology

2011 ◽  
Vol 183-185 ◽  
pp. 259-266
Author(s):  
Xin Sui ◽  
Fu Juan Feng ◽  
Dan Zhao ◽  
Min Min Chen ◽  
Shi Jie Han

A total 408 SSRs were distributed in 18,181 ESTs sequences in Pinaceae in NCBI (National Center for Biotechnology Information) searched by SSRIT software, accounting for 2.24% of the whole EST sequences. We designed 132 pairs of EST-SSR primers by primer3. Of the designed 132 pairs, 29 pairs were able to produce an amplification product in the 10 Pinus koraiensis DNA samples, but only five primers in the 29 pairs exhibited polymorphism. Dinucleotide repeats were the most common repeat class. The repeated primitives of dinucleotide were 10, accounting for 52.73% of the whole repeated primitives; the repeat numbers were 87. The second most common repeat class was trinucleotide. The repeated primitives of trinucleotide were 27, accounting for 42.27% of the whole repeated primitives, and repeat numbers were 78.

Author(s):  
M Idrees ◽  
M Irshad ◽  
ML Pathak ◽  
A Tariq ◽  
R Naeem

The aim of the present study was used to develop SSR markers and to determine genetic relationships among Asparagus species and its cultivars. We mined 72953 Asparagus nucleotide sequences from NCBI and were analyzed. In total, 143 SSRs from 1943 SSRs containing sequences were identified. Of these, 13.6 % were dinucleotide repeats and 2.3 % were trinucleotide repeats. The most frequent dinucleotide repeat motif was AA/TT (73.9 %). The percentage of tri-nucleotide motifs was highest which coded for stop codon (36 %), whereas the Glycine was least present (4.5%). Among the total of 14 SSR primers used, 10 markers yielded 144 (92.3 %) polymorphic bands with an average of 14.4 alleles per primer. Cluster analysis based on UPGMA grouped the Asparagus species and its cultivars into two main clusters. Cluster A contained only A. grecilius which is more diverse than others, while cluster B was further clustered into two sub clusters. Cluster I was comprised of Asparagus officinalis cultivars and cluster II was comprised of Asparagus wild species. Furthermore, these primers (42.8%) were found to be transferable in other medicinal plants (Curcuma longa L. and Saussurea costus). The results suggest that SSR markers are sufficiently useful and powerful to assess genetic relationships and diversity analysis in Asparagus and A. officinalis cultivars. Furthermore, these markers will be particularly useful for evolutionary and genetic mapping studies in Asparagus. J. Biodivers. Conserv. Bioresour. Manag. 2018, 4(2): 21-32


2016 ◽  
Vol 82 (2) ◽  
Author(s):  
Asmini BUDIANI ◽  
Sekar WOELAN ◽  
Hayati MINARSIH ◽  
. NUHAIMI-HARIS ◽  
Riza Arief PUTRANTO

Abstract Breeding program of rubber tree to produce elite clones is hampered by the length of selection cycles. On the other hand, attempts to increase production by extensification of the plantation area is also facing a problem from the availability of the rootstock, causing the occurence of fake clones without any information of their origin. Therefore, the availability of molecular markers to be used as DNA fingerprint of rubber tree clones is needed. This will help the breeder to shorten the length of selection program and to identify the purity of the clone. This research was aimed to evaluate 18 SSR primer pairs that had been published to identify 17 rubber clones. Pure genomic DNAs were isolated from 17 clones, followed by experiment to optimize annealing tempe-rature for each primer to obtain the best amplification product. Initially, the PCR product was run in both the agarose and polyacrylamide gels. However, the analysis of all PCR products were then conducted on SDS polyacrylamide gel, since this gel can separate DNA fragments with only a few bases differences. The results showed that 14 clones have been identified specifically using 11 primers. Four out of 18 primer pairs used could identify 12 rubber tree clones, which are PR 107, PR 261, SP 217, PB 330, PB 340, IRR 5, IRR 112, IRR 118, IRR 220, GT 1, BPM 101 and RRIM 712. Each clone can be distinguished from each other using only one primer pair. Identification of the other tree clones (PB 5/51, PB 260, and RRIC 110) has to be conducted by combining the several PCR products using different primer pairs. Although these results showed that SSR markers had high potential to be used as DNA fingerprint on rubber tree clones, the set of the primer pairs should be tested among other clones, as well as other SSR primers should be tested to identify the clones which could not be identified using the 18 primer pairs in this experiment.Abstrak Pemuliaan tanaman karet untuk menghasilkan klon-klon unggul baru menghadapi masalah lamanya siklus seleksi. Di sisi lain, upaya peningkatan produksi melalui pembukaan lahan baru, juga terkendala oleh ketersediaan bibit, yang memicu beredarnya bibit palsu, yang umumnya tidak jelas asal usulnya. Oleh karena itu, diperlukan ketersediaan marka yang dapat digunakan sebagai sidikjari DNA bagi klon-klon tanaman karet yang ada, sehingga dapat membantu mempercepat proses seleksi dan mengetahui kemurnian bibit. Penelitian ini bertujuan untuk mengevaluasi 18 primer SSR yang telah dipublikasikan untuk mengidentifikasi 17 klon karet. DNA yang murni diisolasi dari 17 klon, kemudian dilakukan optimasi suhu annealing untuk setiap jenis primer agar diperoleh hasil amplifikasi terbaik.  Pada awal percobaan hasil PCR dicek pada gel agarosa dan gel poliakrilamida, namun analisis untuk seluruh hasil PCR dilakukan pada gel SDS poliakrilamid, karena gel ini secara nyata dapat memisahkan fragmen DNA yang hanya berbeda beberapa basa. Hasil percobaan menunjukkan bahwa 14 klon dapat diidentifikasi secara spesifik menggunakan 11 primer. Empat dari 18 pasang primer yang diuji dapat mengidentifikasi 12 klon yang dianalisis, yaitu PR 107, PR 261, SP 217, PB 330, PB 340,. IRR 5, IRR 112, IRR 118, IRR 220, GT 1, BPM 101 dan RRIM 712. Masing-maing klon tersebut dapat dibedakan dari klon lainnya hanya dengan menggunakan satu jenis primer. Sedangkan identifikasi tiga klon lainnya (PB 5/51, PB 260, dan RRIC110) harus dilakukan dengan menggabungkan hasil PCR menggunakan beberapa primer. Meskipun hasil percobaan ini menunjukkan bahwa marka SSR sangat berpotensi untuk digunakan sebagai sidikjari DNA klon-klon karet, namun primer yang sama perlu diuji untuk klon-klon lainnya. Demikian pula primer lain perlu diuji untuk mengidentifikasi klon-klon yang belum teridentifikasi menggunakan 18 primer dalam penelitian ini.


2016 ◽  
Vol 82 (2) ◽  
Author(s):  
Asmini BUDIANI ◽  
Sekar WOELAN ◽  
Hayati MINARSIH ◽  
. NUHAIMI-HARIS ◽  
Riza Arief PUTRANTO

Abstract Breeding program of rubber tree to produce elite clones is hampered by the length of selection cycles. On the other hand, attempts to increase production by extensification of the plantation area is also facing a problem from the availability of the rootstock, causing the occurence of fake clones without any information of their origin. Therefore, the availability of molecular markers to be used as DNA fingerprint of rubber tree clones is needed. This will help the breeder to shorten the length of selection program and to identify the purity of the clone. This research was aimed to evaluate 18 SSR primer pairs that had been published to identify 17 rubber clones. Pure genomic DNAs were isolated from 17 clones, followed by experiment to optimize annealing tempe-rature for each primer to obtain the best amplification product. Initially, the PCR product was run in both the agarose and polyacrylamide gels. However, the analysis of all PCR products were then conducted on SDS polyacrylamide gel, since this gel can separate DNA fragments with only a few bases differences. The results showed that 14 clones have been identified specifically using 11 primers. Four out of 18 primer pairs used could identify 12 rubber tree clones, which are PR 107, PR 261, SP 217, PB 330, PB 340, IRR 5, IRR 112, IRR 118, IRR 220, GT 1, BPM 101 and RRIM 712. Each clone can be distinguished from each other using only one primer pair. Identification of the other tree clones (PB 5/51, PB 260, and RRIC 110) has to be conducted by combining the several PCR products using different primer pairs. Although these results showed that SSR markers had high potential to be used as DNA fingerprint on rubber tree clones, the set of the primer pairs should be tested among other clones, as well as other SSR primers should be tested to identify the clones which could not be identified using the 18 primer pairs in this experiment.Abstrak Pemuliaan tanaman karet untuk menghasilkan klon-klon unggul baru menghadapi masalah lamanya siklus seleksi. Di sisi lain, upaya peningkatan produksi melalui pembukaan lahan baru, juga terkendala oleh ketersediaan bibit, yang memicu beredarnya bibit palsu, yang umumnya tidak jelas asal usulnya. Oleh karena itu, diperlukan ketersediaan marka yang dapat digunakan sebagai sidikjari DNA bagi klon-klon tanaman karet yang ada, sehingga dapat membantu mempercepat proses seleksi dan mengetahui kemurnian bibit. Penelitian ini bertujuan untuk mengevaluasi 18 primer SSR yang telah dipublikasikan untuk mengidentifikasi 17 klon karet. DNA yang murni diisolasi dari 17 klon, kemudian dilakukan optimasi suhu annealing untuk setiap jenis primer agar diperoleh hasil amplifikasi terbaik.  Pada awal percobaan hasil PCR dicek pada gel agarosa dan gel poliakrilamida, namun analisis untuk seluruh hasil PCR dilakukan pada gel SDS poliakrilamid, karena gel ini secara nyata dapat memisahkan fragmen DNA yang hanya berbeda beberapa basa. Hasil percobaan menunjukkan bahwa 14 klon dapat diidentifikasi secara spesifik menggunakan 11 primer. Empat dari 18 pasang primer yang diuji dapat mengidentifikasi 12 klon yang dianalisis, yaitu PR 107, PR 261, SP 217, PB 330, PB 340,. IRR 5, IRR 112, IRR 118, IRR 220, GT 1, BPM 101 dan RRIM 712. Masing-maing klon tersebut dapat dibedakan dari klon lainnya hanya dengan menggunakan satu jenis primer. Sedangkan identifikasi tiga klon lainnya (PB 5/51, PB 260, dan RRIC110) harus dilakukan dengan menggabungkan hasil PCR menggunakan beberapa primer. Meskipun hasil percobaan ini menunjukkan bahwa marka SSR sangat berpotensi untuk digunakan sebagai sidikjari DNA klon-klon karet, namun primer yang sama perlu diuji untuk klon-klon lainnya. Demikian pula primer lain perlu diuji untuk mengidentifikasi klon-klon yang belum teridentifikasi menggunakan 18 primer dalam penelitian ini.


2011 ◽  
Vol 21 (1) ◽  
pp. 18-22
Author(s):  
Rosemary Griffin

National legislation is in place to facilitate reform of the United States health care industry. The Health Care Information Technology and Clinical Health Act (HITECH) offers financial incentives to hospitals, physicians, and individual providers to establish an electronic health record that ultimately will link with the health information technology of other health care systems and providers. The information collected will facilitate patient safety, promote best practice, and track health trends such as smoking and childhood obesity.


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