PMC:4504148 / 11328-12853 JSONTXT

{"project":"2_test","denotations":[{"id":"26236191-9483801-32463215","span":{"begin":142,"end":146},"obj":"9483801"},{"id":"26236191-12853124-32463216","span":{"begin":263,"end":267},"obj":"12853124"},{"id":"26236191-1561104-32463217","span":{"begin":939,"end":943},"obj":"1561104"},{"id":"26236191-12853124-32463220","span":{"begin":263,"end":267},"obj":"12853124"}],"text":"Yeast strains and growth conditions\nIn this study, we used the BY4741 (MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0) wild-type strain (Brachmann et al., 1998). Full-length cDNA of either wtDFNA5 or mutDFNA5 was isolated and amplified as previously described (Gregan et al., 2003). Amplified products were ligated into yeast pYX212 plasmid containing an HA-marker (Clontech, Mountain View, CA, USA) using EcoRI and BamHI restriction sites. All constructs were verified by bidirectional sequencing on an ABI genetic analyser 3130 × l (AppliedBiosystems, FosterCity, CA, USA).\nYeast strains were grown at 30°C in selective medium containing 2% glucose (SD-URA). Fifty milliliter yeast cultures, transformed with either wtDFNA5 or mutDFNA5, were harvested in mid-exponential phase (OD600 nm = 3.5−4.2) and at the post-diauxic shift (OD600 nm = 7.4−8.4) (Figure 1). Standard transformation techniques were applied for these transformations (Gietz et al., 1992).\nFigure 1 Yeast microarray design. (A) Illustration of the different comparisons that were made between the RNA samples. RNA was collected from yeast strains transformed with either wtDFNA5 or mutDFNA5 at two different time-points. The bold and dashed lines represent the color flip of each RNA sample. (B) Growth profile of the Saccharomyces cerevisiae BY4741 background strain transformed with either wtDFNA5 (gray squares) or mutDFNA5 (black squares). The blue and red ellipse illustrate respectively the mid-exponential and post-diauxic time-points when RNA was collected.\n\nR"}

{"project":"MyTest","denotations":[{"id":"26236191-9483801-32463215","span":{"begin":142,"end":146},"obj":"9483801"},{"id":"26236191-12853124-32463216","span":{"begin":263,"end":267},"obj":"12853124"},{"id":"26236191-1561104-32463217","span":{"begin":939,"end":943},"obj":"1561104"},{"id":"26236191-12853124-32463220","span":{"begin":263,"end":267},"obj":"12853124"}],"namespaces":[{"prefix":"_base","uri":"https://www.uniprot.org/uniprot/testbase"},{"prefix":"UniProtKB","uri":"https://www.uniprot.org/uniprot/"},{"prefix":"uniprot","uri":"https://www.uniprot.org/uniprotkb/"}],"text":"Yeast strains and growth conditions\nIn this study, we used the BY4741 (MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0) wild-type strain (Brachmann et al., 1998). Full-length cDNA of either wtDFNA5 or mutDFNA5 was isolated and amplified as previously described (Gregan et al., 2003). Amplified products were ligated into yeast pYX212 plasmid containing an HA-marker (Clontech, Mountain View, CA, USA) using EcoRI and BamHI restriction sites. All constructs were verified by bidirectional sequencing on an ABI genetic analyser 3130 × l (AppliedBiosystems, FosterCity, CA, USA).\nYeast strains were grown at 30°C in selective medium containing 2% glucose (SD-URA). Fifty milliliter yeast cultures, transformed with either wtDFNA5 or mutDFNA5, were harvested in mid-exponential phase (OD600 nm = 3.5−4.2) and at the post-diauxic shift (OD600 nm = 7.4−8.4) (Figure 1). Standard transformation techniques were applied for these transformations (Gietz et al., 1992).\nFigure 1 Yeast microarray design. (A) Illustration of the different comparisons that were made between the RNA samples. RNA was collected from yeast strains transformed with either wtDFNA5 or mutDFNA5 at two different time-points. The bold and dashed lines represent the color flip of each RNA sample. (B) Growth profile of the Saccharomyces cerevisiae BY4741 background strain transformed with either wtDFNA5 (gray squares) or mutDFNA5 (black squares). The blue and red ellipse illustrate respectively the mid-exponential and post-diauxic time-points when RNA was collected.\n\nR"}