diff --git a/assays/plant_genotyping/dataset/complementation_F1_genotype_results.xlsx b/assays/plant_genotyping/dataset/complementation_F1_genotype_results.xlsx index de90a5718db7c4e8bf61df8b846ade199fce3aa6..b0272142474b373941fb149530cc30ea13ce1bde 100644 Binary files a/assays/plant_genotyping/dataset/complementation_F1_genotype_results.xlsx and b/assays/plant_genotyping/dataset/complementation_F1_genotype_results.xlsx differ diff --git a/assays/plant_genotyping/protocols/complementation_plant_genotyping.md b/assays/plant_genotyping/protocols/complementation_plant_genotyping.md new file mode 100644 index 0000000000000000000000000000000000000000..54ea91ec91beb89bc95ccfea5117d0c158ac8be6 --- /dev/null +++ b/assays/plant_genotyping/protocols/complementation_plant_genotyping.md @@ -0,0 +1,11 @@ +**F1 plants of lwd/eam7 allelism test: genoypting ** + +- DNA was extracted from leaves of all plants using QIAGEN_DNeasy-Plant_mini protocol +- plants were tested for possible mutations in the CDS of LWD1 by amplifying the full genomic sequence using the primers LWD1_71us_F (5'-ATCAGACCCCTCCAACGACT-3') and LWD1_21ds_R (5'-TTCCTCTTTCTACCCCACCA-3') +- the PCR fragments were Sanger sequenced using two different primers: + - LWD1_71us_F (test the beginning of the CDS) + - LWD1_qPCR114F (5'-CAGCTCATAGCCCATGACAA-3') (test the end of the CDS) +- all mutations and the zygosity of the mutations were determined relative to the start codon of LWD1 + +- all F1 plants were genotyped +- only some parent plants were genotyped again \ No newline at end of file