restriction enzymes bstapi  (New England Biolabs)

Journal: Chromosoma

Article Title: Establishment and inheritance of minichromosomes from Arabidopsis haploid induction

doi: 10.1007/s00412-023-00788-5

Figure Lengend Snippet: Primer sequences used for mini1a and mini3b detection

Article Snippet: After PCR, a 5 μL digestion mix containing 0.1 μL restriction enzyme SspI (for SNP1:13,405,811) or BstAPI (for SNP3:16,409,164), 1 μL CutSmart Buffer (NEB), and 3.9 uL nuclease-free water was added to final PCR and digested overnight at 37 °C.

Techniques: Sequencing

Journal: Chromosoma

Article Title: Establishment and inheritance of minichromosomes from Arabidopsis haploid induction

doi: 10.1007/s00412-023-00788-5

Figure Lengend Snippet: Detection and analysis of minichromosomes. The presence of mini1a and mini3b minichromosomes derived from the GFP-tailswap haploid inducer (Col-0 ecotype) in haploids induced from a Nfa8/Sq8 F1 hybrid was confirmed by genomic analysis. A , B Detection of Col-0 SNPs on chromosome 1 (Chr1) and chromosome 3 (Chr3) of haploid individuals. C , D Dosage plot of F2 doubled haploid lines containing mini1a and mini3b . E Origin, inferred structure, and breakpoint junction of mini1a at circularization site. F A cleaved amplified polymorphic sequence (CAPS) assay using the restriction enzyme SspI to distinguish Sq-8/NFA-8 lines containing mini1a using a SNP at position 13,405,811 of Chr1, as well as corresponding mini1a junction PCR of mini1a at the expected breakpoint junction site

Article Snippet: After PCR, a 5 μL digestion mix containing 0.1 μL restriction enzyme SspI (for SNP1:13,405,811) or BstAPI (for SNP3:16,409,164), 1 μL CutSmart Buffer (NEB), and 3.9 uL nuclease-free water was added to final PCR and digested overnight at 37 °C.

Techniques: Derivative Assay, Amplification, Sequencing