Purification:Article Title: The genomic history of Iberian horses since the last Ice Age
Article Snippet: .. The end-repaired DNA templates were subsequently purified using MinElute columns, as described above, and then ligated with Quick T4 Ligase Enzyme (NEBNext® Quick Ligation Module, NEB E6056, 2.5 μL in a final reaction volume of 25 μL) to DNA adapters comprising the Illumina DNA sequencing primer followed by a unique 7-bp index93. .. Following purification, DNA libraries underwent a filling-in step using Bst DNA Polymerase (NEB M0275, final concentration = 0.48 U/μL) for 20 minutes at 37oC, followed by inactivation for 20 minutes at 80°C.
Ligation:Article Title: The genomic history of Iberian horses since the last Ice Age
Article Snippet: .. The end-repaired DNA templates were subsequently purified using MinElute columns, as described above, and then ligated with Quick T4 Ligase Enzyme (NEBNext® Quick Ligation Module, NEB E6056, 2.5 μL in a final reaction volume of 25 μL) to DNA adapters comprising the Illumina DNA sequencing primer followed by a unique 7-bp index93. .. Following purification, DNA libraries underwent a filling-in step using Bst DNA Polymerase (NEB M0275, final concentration = 0.48 U/μL) for 20 minutes at 37oC, followed by inactivation for 20 minutes at 80°C.
DNA Sequencing:Article Title: The genomic history of Iberian horses since the last Ice Age
Article Snippet: .. The end-repaired DNA templates were subsequently purified using MinElute columns, as described above, and then ligated with Quick T4 Ligase Enzyme (NEBNext® Quick Ligation Module, NEB E6056, 2.5 μL in a final reaction volume of 25 μL) to DNA adapters comprising the Illumina DNA sequencing primer followed by a unique 7-bp index93. .. Following purification, DNA libraries underwent a filling-in step using Bst DNA Polymerase (NEB M0275, final concentration = 0.48 U/μL) for 20 minutes at 37oC, followed by inactivation for 20 minutes at 80°C.
Clone Assay:Article Title: Heat-Resistant Agglutinin 1 Is an Accessory Enteroaggregative Escherichia coli Colonization Factor
Article Snippet: .. Unless otherwise stated, ligations were performed using Quick T4 ligase enzyme (New England Biolabs), and clones and plasmids were transformed into chemically competent E. coli K-12 DH5α or TOP10 cells. .. Transformation into EAEC strains was accomplished by electroporation using 2-mm cuvettes with a Bio-Rad Micropulser, according to the manufacturer's instructions. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Primer Purpose Sequence Reference PUC18KNcoIF Amplification of apha -3 cassette and attachment of NcoI tails 5′-CATGCCATGGTGACTAAACTAGGAGGAATAAATGGCTAAAATGAGA-3′ This study PUC18KNcoIR Amplification of apha -3 cassette and attachment of NcoI tails 5′-CATGCCATGGTCATTATTCCCTCCAGGTACTAAAACAATTCATC-3′ This study Agg1F Primer internal to hra1 ; used to verify mutant 5′-ATTGCGGTTTCAGCGCTTGC-3′ This study Agg1R Primer internal to hra1 ; used to verify mutant 5′-AGATAGCGATAGCTGAGGTC-3′ This study Hra1upF Upstream of hra1 but absent from hra1 deletion construct pSB1; used for primary cloning of the gene and to verify mutant 5′-GCAAATCACAACCTTCCTGA-3′ This study Hra1dnR Downstream of hra1 and present in hra1 deletion construct pSB1; used for primary cloning of gene and to verify mutant 5′-CTTCGGAGTGGATGTATCTG-3′ This study AA432F Amplifies portion of aggregative plasmid gene aatA ; used to verify mutant 5′-CTGGCGAAAGACTGTATCAT-3′ 8 AA432R Amplifies portion of aggregative plasmid gene aatA ; used to verify mutant 5′-CAATGTATAGAAATCCGCTGTT-3′ 8 Open in a separate window Oligonucleotide primers used in this study Cloning the hra1 gene from EAEC strain 042.
Article Title: Error-Prone DNA Repair System in Enteroaggregative Escherichia coli Identified by Subtractive Hybridization
Article Snippet: .. Unless otherwise stated, ligations were performed by using Quick T4 ligase enzyme (NEB), and clones and plasmids were transformed into chemically competent E. coli K-12 DH5 or TOP10 cells. .. Transformation of large plasmids into Electromax DH5 -E cells (Invitrogen) and all plasmids into EAEC strains was accomplished by electroporation using a Micropulser (Bio-Rad) according to the manufacturer’s instructions.
Transformation Assay:Article Title: Heat-Resistant Agglutinin 1 Is an Accessory Enteroaggregative Escherichia coli Colonization Factor
Article Snippet: .. Unless otherwise stated, ligations were performed using Quick T4 ligase enzyme (New England Biolabs), and clones and plasmids were transformed into chemically competent E. coli K-12 DH5α or TOP10 cells. .. Transformation into EAEC strains was accomplished by electroporation using 2-mm cuvettes with a Bio-Rad Micropulser, according to the manufacturer's instructions. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Primer Purpose Sequence Reference PUC18KNcoIF Amplification of apha -3 cassette and attachment of NcoI tails 5′-CATGCCATGGTGACTAAACTAGGAGGAATAAATGGCTAAAATGAGA-3′ This study PUC18KNcoIR Amplification of apha -3 cassette and attachment of NcoI tails 5′-CATGCCATGGTCATTATTCCCTCCAGGTACTAAAACAATTCATC-3′ This study Agg1F Primer internal to hra1 ; used to verify mutant 5′-ATTGCGGTTTCAGCGCTTGC-3′ This study Agg1R Primer internal to hra1 ; used to verify mutant 5′-AGATAGCGATAGCTGAGGTC-3′ This study Hra1upF Upstream of hra1 but absent from hra1 deletion construct pSB1; used for primary cloning of the gene and to verify mutant 5′-GCAAATCACAACCTTCCTGA-3′ This study Hra1dnR Downstream of hra1 and present in hra1 deletion construct pSB1; used for primary cloning of gene and to verify mutant 5′-CTTCGGAGTGGATGTATCTG-3′ This study AA432F Amplifies portion of aggregative plasmid gene aatA ; used to verify mutant 5′-CTGGCGAAAGACTGTATCAT-3′ 8 AA432R Amplifies portion of aggregative plasmid gene aatA ; used to verify mutant 5′-CAATGTATAGAAATCCGCTGTT-3′ 8 Open in a separate window Oligonucleotide primers used in this study Cloning the hra1 gene from EAEC strain 042.
Article Title: Error-Prone DNA Repair System in Enteroaggregative Escherichia coli Identified by Subtractive Hybridization
Article Snippet: .. Unless otherwise stated, ligations were performed by using Quick T4 ligase enzyme (NEB), and clones and plasmids were transformed into chemically competent E. coli K-12 DH5 or TOP10 cells. .. Transformation of large plasmids into Electromax DH5 -E cells (Invitrogen) and all plasmids into EAEC strains was accomplished by electroporation using a Micropulser (Bio-Rad) according to the manufacturer’s instructions.
Article Title: The Heat-Resistant Agglutinin Family Includes a Novel Adhesin from Enteroaggregative Escherichia coli Strain 60A
Article Snippet: .. Unless otherwise stated, ligations were performed using Quick T4 ligase enzyme (New England BioLabs), and plasmids were transformed into chemically competent E. coli K-12 TOP10 cells. .. Transformation of plasmids into EAEC strains was accomplished by electroporation using a Micropulser (Bio-Rad) according to the manufacturer’s instructions.
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