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Found 3 matching solutions for this experiment
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pGK43-SEAP and pGK73-SAEP were linearized by digestion with BstZ17I and SgrAI, whereas genomic plasmids pAd4ΔE3 or pAd7ΔE3 were linearized by digestion with SwaI (Thermo Fisher Scientific, Waltham, MA, United States), and pAd4-SEAP and pAd7-SEAP were constructed by homologous recombination between linearized shuttle reporter plasmids and genomic plasmids. |
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pGK43-SEAP and pGK73-SAEP were linearized by digestion with BstZ17I and SgrAI, whereas genomic plasmids pAd4ΔE3 or pAd7ΔE3 were linearized by digestion with SwaI (Thermo Fisher Scientific, Waltham, MA, United States), and pAd4-SEAP and pAd7-SEAP were constructed by homologous recombination between linearized shuttle reporter plasmids and genomic plasmids. |
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For direct assembly of transformable plasmids by cycled ligation, a modified pUC19 vector (Table S1) with two blunt ligation sites separated by ∼250 bp was created. The vector was digested with NruI and SwaI (NEB) and the linearized vector was isolated and purified by gel extraction (Qiagen). Detailed protocol in Table S3. |
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For direct assembly of transformable plasmids by cycled ligation, a modified pUC19 vector (Table S1) with two blunt ligation sites separated by ∼250 bp was created. The vector was digested with NruI and SwaI (NEB) and the linearized vector was isolated and purified by gel extraction (Qiagen). Detailed protocol in Table S3. |
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To estimate the genome size, DNA was digested with two rare-cutting restriction enzymes, PmeI (NEB, Ipswich, MA) and SmiI (recognition site identical to that of SwaI) (TaKaRa). |
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To estimate the genome size, DNA was digested with two rare-cutting restriction enzymes, PmeI (NEB, Ipswich, MA) and SmiI (recognition site identical to that of SwaI) (TaKaRa). |
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