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Our Case
  • Screening homozygous transgenic plants by flanking

    Traditional method of screening homozygous transgenic plants is several generations self-pollenation, it is a time consuming work and low efficiency. A new method by flanking sequences amplification of T-DNA insertion site to screen homozygous plants was created.

  • A simple and cost-effective method for screening of

    2018-5-29 · The main advantages of our method are its technical simplicity, quickness and low cost, it could be used to screen homozygous/biallelic mutants from T 0 plants, or the descendants of heterozygous/monoalleic mutants. Moreover, the two-round of PCR enhanced the accuracy and reproducibility of the PCR results.

  • Fast-tracking development of homozygous transgenic

    2013-4-30 · To obtain homozygous lines carrying a unique insertion event as efficiently as possible, T0 plants (plants raised from transformed callus) with a single copy of the transgene were selected and their progeny screened for homozygous plants. Finally, the assay was adapted to work on rice.

  • Arabidopsis crossing and how to select homozygous

    The simple way is identifying plants that have both genes (PCR, selection agent...). Grown these plants to have the next generation and screen again to find if they have segregation of the two genes.

  • A Genetic Screen for Nitrate Regulatory Mutants

    YFP expression was induced by nitrate in these plants. Homozygous transgenic plants were then ethyl methanesulfonate mutagenized to produce M2 seed-lings, of which approximately 35,000 were screened for low YFP fluorescence after nitrate treatment. Ini-tially

  • A CRISPR‐Cas9‐mediated domain‐specific base‐editing

    generation, T-DNA-free homozygous I1879V plants and heterozy-gous C2186R plants were isolated and sprayed with a commercial APP herbicide Gallant to confirm resistance and evaluate the potential usage of mutations in fields. Wild-type (WT) plants died after 7 days, while all mutants demonstrated significant resistance (Figure 1e). Compared with WT, no obvious differences were

  • A simple and efficient method for CRISPR/Cas9

    2017-4-20 · Here, we selected the T 1 generation of these plants to screen for homozygous mutants using ACT-PCR. The critical annealing temperatures of both OsLG1 and OsGL1-1 primers were found to be 66.7°C (Fig. 3A and B). Subsequent PCR showed that amplifications were disrupted in 5 of the 24 OsLG1 T 1 plants and 7 of the 36 OsGL1-1 T 1 plants (Fig. 3C and D

  • High‐throughput detection and screening of plants

    2018-5-15 · In total, 15 T 0 transgenic plants, including Arabidopsis thaliana, Sorghum bicolor, and Zea mays (five samples/species), were used to screen out positive gene‐edited individuals. Figure 7 (a) shows that the target gene AT5G05570 in Arabidopsis thaliana was detected.

  • Regulation of plant immune receptor accumulation

    2017-3-31 · For fine mapping, about five hundred F3 plants generated from F2 plants heterozygous for muse11, wild-type for mos2 or mos4, and homozygous for snc1 were used. After the muse11 mutation was narrowed down between markers MUL8 (15.7 Mb) and K23L20 (18.02 Mb) on Chromosome 5, next-generation re-sequencing was performed to identify mutations within the muse11 region.

  • EMS Mutagenesis of Arabidopsis Seed CSH Protocols

    INTRODUCTION. EMS-mutagenized Arabidopsis seed collections can be purchased from companies or acquired from other researchers. But in some cases (e.g., mutagenesis of a specific genotype), it is necessary for an investigator to generate EMS-mutagenized seed.

  • Frontiers An Efficient Visual Screen for CRISPR/Cas9

    2017-1-24 · Homozygous T-DNA insertion plants were identified using the gene specific primer pair FH214 and FH215 (Supplementary Table S1) as well as the T-DNA specific primer pair FH215 and P49 (Supplementary Table S1). Homozygosity was additionally confirmed by visual control of trichome loss. Homozygous gl1 plants were further propagated.

  • Using plants for biofuel production Stanford University

    2014-11-19 · M1 plants (heterozygous) M2 seed . families . M2 populations (segregating ¼ homozygous) M3 seed EMS mutagenesis Identify mutant gene . Further detailed characterisation Simple visual screen for plants with restored growth Suppressor screen

  • Frontiers The HEM Lines: A New Library of

    2018-9-19 · Whole genome sequencing of a subset of lines showed an average of 696 homozygous mutations per line, 195 of which (28%) modify a protein sequence. To test the power of this library, we carried out a forward screen looking for meiotic defects by observing chromosomes at

  • A simple and efficient method for CRISPR/Cas9

    2017-4-20 · Here, we selected the T 1 generation of these plants to screen for homozygous mutants using ACT-PCR. The critical annealing temperatures of both OsLG1 and OsGL1-1 primers were found to be 66.7°C (Fig. 3A and B). Subsequent PCR showed that amplifications were disrupted in 5 of the 24 OsLG1 T 1 plants and 7 of the 36 OsGL1-1 T 1 plants (Fig. 3C

  • Genetic evidence for the in planta role of phloem

    In the homozygous state, these mutations resulted in stunted growth, retarded development, and sterility. The source leaves of mutant plants contained a great excess of starch, and radiolabeled sugar failed to be transported efficiently to roots and inflorescences.

  • WO 2013/103366 A1 A Method To Screen Plants For

    A METHOD TO SCREEN PLANTS FOR GENETIC ELEMENTS INDUCING . PARTHENOGENESIS IN PLANTS . FIELD OF THE DISCLOSURE . The present disclosure relates to the field of plant molecular biology, more particularly to plant female reproductive biology, methods of altering plant female reproductive biology and screening for altered mechanistic capacities for reproduction.

  • Mutagenomics: A Rapid, High-Throughput Method to

    Genetic screens are powerful tools to dissect complex biological processes, but a rate-limiting step is often the cloning of targeted genes. Here, we present a strategy, “mutagenomics,” to identify causal mutations from a screen in a high throughput fashion in the absence of backcrossing. Mutagenomics is initiated by sequencing the genomes of the mutants identified, which are then

  • Regulation of plant immune receptor accumulation

    2017-3-31 · For fine mapping, about five hundred F3 plants generated from F2 plants heterozygous for muse11, wild-type for mos2 or mos4, and homozygous for snc1 were used. After the muse11 mutation was narrowed down between markers MUL8 (15.7 Mb) and K23L20 (18.02 Mb) on Chromosome 5, next-generation re-sequencing was performed to identify mutations within

  • T-DNA as an Insertional Mutagen in Arabidopsis

    1999-12-1 · Mutations that are homozygous lethal can be maintained in the population in the form of heterozygous plants. Polymerase chain reaction (PCR) methods have been developed that allow one to easily isolate individual plants that carry a particular T-DNA mutation of interest (McKinney et al., 1995; Krysan et al., 1996).

  • EMS Mutagenesis of Arabidopsis Seed CSH Protocols

    INTRODUCTION. EMS-mutagenized Arabidopsis seed collections can be purchased from companies or acquired from other researchers. But in some cases (e.g., mutagenesis of a specific genotype), it is necessary for an investigator to generate EMS-mutagenized seed.