Journal: Gene

Article Title: Enrichment of meiotic recombination hotspot sequences by avidin capture technology 2

doi: 10.1016/j.gene.2012.12.042

Figure Lengend Snippet: Principle of the capture protocol. The hybridization of oligos 1 and 2 creates an EcoRI restriction site and a 5’ end overhang that is complementary to the sequence in the recombination hotspot 5’-CCN CCN TNN CCN C-3’. The annealed capture oligonucleotides are immobilized on streptavidin-magnetic coated beads through the biotin-conjugated 5’ end in oligo 1.

Article Snippet: Oligos were denatured by heating at 95°C for 5 min in a thermocycler, followed by annealing at 22°C for 1 h. The annealed oligos have an EcoRI restriction site ( ).

Techniques: Hybridization, Sequencing

Journal: Gene

Article Title: Enrichment of meiotic recombination hotspot sequences by avidin capture technology 2

doi: 10.1016/j.gene.2012.12.042

Figure Lengend Snippet: Enrichment of a DNA containing the hotspot motif from a mixture of short, double-stranded DNA. (A) Synthetic dsDNA oligonucleotides: 100-mer containing the hotspot sequence for recombination; 50-mer not containing the hotspot sequence. (B) PCR-amplified samples of supernatants from streptavidin beads after loading with oligonucleotides (sample 1), sequential washes of beads (samples 2 – 9), and supernatant after treating beads with EcoRI (sample 10). (C) As described for “A” but now the 50-mer contains the hotspot sequence. (D) As described for “B.” Abbreviation: M, marker.

Article Snippet: Oligos were denatured by heating at 95°C for 5 min in a thermocycler, followed by annealing at 22°C for 1 h. The annealed oligos have an EcoRI restriction site ( ).

Techniques: Sequencing, Amplification, Marker

Journal: Gene

Article Title: Enrichment of meiotic recombination hotspot sequences by avidin capture technology 2

doi: 10.1016/j.gene.2012.12.042

Figure Lengend Snippet: Capture of long dsDNA with oligo-Blue. (A) EcoRI fragment of pBluescript II sk (+), containing a 13-mer with a degree of degeneration similar to the hotspot sequence. (B) PCR-amplified samples of supernatants from streptavidin beads after loading with oligonucleotides (sample 1), sequential washes of beads (samples 2 – 9), and supernatant after treating beads with EcoRI (sample 10). Abbreviation: M, marker.

Article Snippet: Oligos were denatured by heating at 95°C for 5 min in a thermocycler, followed by annealing at 22°C for 1 h. The annealed oligos have an EcoRI restriction site ( ).

Techniques: Sequencing, Amplification, Marker