Young trifoliate leaf tissue was collected from the first replicate block of each population at the Palmyra 2016 location. Leaf tissue for each RIL was sampled from multiple plants in each plot and stored in 2 mL screw cap tubes. The samples were freeze-dried for 72-h using a Savant ModulyoD Thermoquest (Savant Instruments, Holbrook, NY), and then stored at − 80 °C for future use. Genomic DNA was extracted from the freeze-dried tissue samples using a modified procedure from the Sigma GenElute™ DNA Extraction Kit (SIGMA®, Saint Louis, MO) methodology. DNA quality was verified using electrophoresis with 1% agarose gels, while quantity was verified using a Qubit® 2.0 fluorometer (Invitrogen, Carlsbad, CA). DNA samples (30 μl of 10 ng μl− 1 DNA) were transferred to Plate-forme D’analyses Génomiques at Université Laval (Laval, Quebec, Canada) for genotyping-by-sequencing (GBS), using the Fast-GBS pipeline with the Gmax_275_v2 reference genome [88 (link)]. The Fast-GBS pipeline identified 24,738 high-quality single-nucleotide polymorphisms (SNPs). Heterozygous SNPs were considered missing data. SNPs with > 20% missing data or a minimum minor allele frequency less than 0.3 were discarded prior to imputation with Beagle [89 (link)].
Sigma genelute dna extraction kit
Manufactured by Merck Group
The Sigma GenElute™ DNA Extraction Kit is a product designed for the purification of DNA from a variety of sample types. It utilizes a silica-based membrane technology to efficiently capture and purify DNA, which can then be eluted for further analysis or applications.
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Lab products found in correlation
2 protocols using sigma genelute dna extraction kit
1
Genetic Variation Analysis of Trifoliate Leaves
2
High-Quality DNA Extraction and GBS Genotyping
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Young trifoliate leaf tissue was collected from the first replicate block of each population at the Palmyra 2016 location. Leaf tissue for each RIL was sampled from multiple plants in each plot and stored in 2mL screw cap tubes. The samples were freeze-dried for 72-hours using a Savant ModulyoDThermoquest (Savant Instruments, Holbrook, NY), and then stored at -80°C for future use. Genomic DNA was extracted from the freeze-dried tissue samples using a modified procedure from the Sigma GenElute™ DNA Extraction Kit (SIGMA®, Saint Louis, MO) methodology. DNA quality was verified using electrophoresis with 1% agarose gels, while quantity was verified using a Qubit ® 2.0 fluorometer (Invitrogen, Carlsbad, CA).
DNA samples (30μl of 10ng μl -1 DNA) were transferred to Plate-formeD'analysesGénomiques at Université Laval (Laval, Quebec, Canada) for genotyping-by-sequencing (GBS), using the Fast-GBS pipeline with the Gmax_275_v2 reference genome [75] . The Fast-GBS pipeline identified 24,738 high-quality single-nucleotide polymorphisms (SNPs). Heterozygous SNPs were considered missing data. SNPs with >20% missing data or a minimum minor allele frequency less than 0.3 were discarded prior to imputation with Beagle [76] .
DNA samples (30μl of 10ng μl -1 DNA) were transferred to Plate-formeD'analysesGénomiques at Université Laval (Laval, Quebec, Canada) for genotyping-by-sequencing (GBS), using the Fast-GBS pipeline with the Gmax_275_v2 reference genome [75] . The Fast-GBS pipeline identified 24,738 high-quality single-nucleotide polymorphisms (SNPs). Heterozygous SNPs were considered missing data. SNPs with >20% missing data or a minimum minor allele frequency less than 0.3 were discarded prior to imputation with Beagle [76] .
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