diff --git a/assays/plant_genotyping/protocols/lwd1_plant_genotyping.md b/assays/plant_genotyping/protocols/lwd1_plant_genotyping.md index 0a4205e105c40440930cdec0dcdb202fe506cf04..be2bdbe65765f52c27ca081dc6d019f550f82f10 100644 --- a/assays/plant_genotyping/protocols/lwd1_plant_genotyping.md +++ b/assays/plant_genotyping/protocols/lwd1_plant_genotyping.md @@ -1,4 +1,6 @@ -**lwd1 T0 and T1 plants ** +## Genotyping of *lwd1* mutants + +***lwd1* T0 and T1 plants:** - DNA was extracted from leaves of all mutants 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') @@ -7,5 +9,5 @@ - plants transformed with pMGE599/182-1222 (approach 2): LWD1_qPCR114F (5'-CAGCTCATAGCCCATGACAA-3') - mutations and the zygosity of the mutations were determined relative to the start codon of LWD1 -**only in T1 plants**: +**Only in T1 plants:**: - PCR was performed on transformation vector to check presence/absence of Cas9 using the primers pMGE599_M13F (5'-GTTTTCCCAGTCACGAC-3') and pMGE599_R (5'-TGTCGACATCGATAAGCTTGAA-3') \ No newline at end of file