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The selected candidate was identified by sequencing the 16S rRNA.
The underlying causative gene, UBIAD1, was identified by sequencing genes of the interval [17].
The calcium and integrin binding protein CIB1 was identified by sequencing and the interaction was confirmed by immunoprecipitation experiments.
Although many of the original alleles were not available for these studies, a stop codon was identified by sequencing the spg242 (previously called spg2) allele (W487*).
Although one small (26 bp) LKB1 deletion representing a complete loss-of-function was identified by sequencing (Table S1, Figure 1A), larger deletions would have been missed.
After the recombinant was identified by sequencing, pAdTrack-CMV/pGEM T-Easy-P311 shuttle plasmid was constructed and transfected into competent E. coli DH5α (ATCC, USA), followed by collection of rigid clones.
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Positive transformants were identified by sequencing (Baseclear, Leiden, The Netherlands).
UL54 and UL97 gene mutations were identified by sequencing.
Bacteria that showed AMP-degrading activity were identified by sequencing the 16S rRNA gene.
ORFs determined to express soluble recombinant proteins were identified by sequencing and analysed by small-scale IMAC and SDS PAGE.
Over forty such alleles have been identified by sequencing the KIT genes of various horses.
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