A High Through-Put Protocol of Plant Genomic DNA Preparation for PCR

Zhang, Guohua; Gao, Mingyang; Zhang, Guizhi; Sun, Jin-jie; Jin, Xue-Mei; Wang, Chunyang; Zhao, Yan; Li, Sishen

doi:10.3724/sp.j.1006.2013.01200

Cited by 12 publications

(2 citation statements)

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“…Genomic DNA was extracted from fresh rice leaves following the high-throughput protocol of plant genomic DNA preparation (Wang et al 2013). PCR reactions were performed using a 96-well plate with a 10-μL reaction mixture that contained approximately 50 ng genomic DNA, 1 μL 10× Easy Taq buffer (Transgen Biotech Inc., Beijing, China), 0.2 μL 2.5 mM dNTPs (Transgen Biotech Inc., Beijing, China), 0.5 U Easy Taq DNA Polymerase (Transgen Biotech Inc., Beijing, China), and 0.125 μL 20× EvaGreen (Biotium Inc., USA); the reactions were covered with 10 μL mineral oil (Amresco Inc., USA).…”

Section: Methodsmentioning

confidence: 99%

Updating the elite rice variety Kongyu 131 by improving the Gn1a locus

Feng

Wang

Nan

et al. 2017

Rice

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BackgroundKongyu 131 is an elite japonica rice variety of Heilongjiang Province, China. It has the characteristics of early maturity, superior quality, high yield, cold tolerance and wide adaptability. However, there is potential to improve the yield of Kongyu 131 because of the relatively few grains per panicle compared with other varieties. Hence, we rebuilt the genome of Kongyu 131 by replacing the GRAIN NUMBER1a (Gn1a) locus with a high-yielding allele from a big panicle indica rice variety, GKBR. High-resolution melting (HRM) analysis was used for single nucleotide polymorphism (SNP) genotyping.ResultsQuantitative trait locus (QTL) analysis of the BC3F2 population showed that the introgressed segment carrying the Gn1a allele of GKBR significantly increased the branch number and grain number per panicle. Using 5 SNP markers designed against the sequence within and around Gn1a, the introgressed chromosome segment was shortened to approximately 430 Kb to minimize the linkage drag by screening recombinants in the target region. Genomic components of the new Kongyu 131 were detected using 220 SNP markers evenly distributed across 12 chromosomes, suggesting that the recovery ratio of the recurrent parent genome (RRPG) was 99.89%. Compared with Kongyu 131, the yield per plant of the new Kongyu 131 increased by 8.3% and 11.9% at Changchun and Jiamusi, respectively.ConclusionsTo achieve the high yield potential of Kongyu 131, a minute chromosome fragment carrying the favorable Gn1a allele from the donor parent was introgressed into the genome of Kongyu 131, which resulted in a larger panicle and subsequent yield increase in the new Kongyu 131. These results indicate the feasibility of improving an undesirable trait of an elite variety by replacing only a small chromosome segment carrying a favorable allele.

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Section: Methodsmentioning

confidence: 99%

Updating the elite rice variety Kongyu 131 by improving the Gn1a locus

Feng

Wang

Nan

et al. 2017

Rice

View full text Add to dashboard Cite

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“…In the case of large-scale genotyping with DNA markers, e.g., SSRs (simple sequence repeats), indels (insertions/deletions), and SNPs (single nucleotide polymorphisms), preparation of gDNA templates from massive samples is time-consuming and tedious. Therefore, a very high-throughput gDNA preparation method has been developed to deal with large number of samples for PCR-based genotyping (Wang et al, 2013). Very-large-size gDNA is necessary for large-insert genomic library construction and long-range physical mapping.…”

Section: Introductionmentioning

confidence: 99%

Preparation of Rice Plant Genomic DNA for Various Applications

Zhou

Shen

et al. 2016

CP Plant Biology

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Research on plant molecular biology, genetic engineering, and crop molecular breeding involves various manipulations of plant genomic DNAs (gDNAs). These studies require preparing gDNAs from plant materials using suitable methods according to the yield, intactness, and purity of the resultant gDNA samples. Here we describe protocols for preparation of rice plant gDNAs for various applications including: (1) maxipreps and (2) minipreps of gDNAs for restriction analysis, gene cloning, PCR amplification, genomic sequencing, and genotyping; (3) preparation of megabase‐sized nuclear DNA for construction of large‐insert genomic libraries and long‐range physical mapping; and (4) 96‐well‐format high‐throughput gDNA preparation for PCR‐based genotyping. The methods are also suitable for other plant species including dicots. © 2016 by John Wiley & Sons, Inc.

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Development and utilization of functional KASP markers to improve rice eating and cooking quality through MAS breeding

Yang

Chen

et al. 2019

Euphytica

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A High Through-Put Protocol of Plant Genomic DNA Preparation for PCR

Cited by 12 publications

References 0 publications

Updating the elite rice variety Kongyu 131 by improving the Gn1a locus

Updating the elite rice variety Kongyu 131 by improving the Gn1a locus

Preparation of Rice Plant Genomic DNA for Various Applications

Development and utilization of functional KASP markers to improve rice eating and cooking quality through MAS breeding

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