Polymerase Chain Reaction:Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription.
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [19] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [ ] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Amplification:Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription.
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [19] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [ ] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Introduce:Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription.
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [19] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [ ] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Clone Assay:Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription.
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [19] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [ ] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Generated:Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription.
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [19] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [ ] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Mutagenesis:Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription.
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [19] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
Article Title: The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription
Article Snippet: All plasmids used in this study have been deposited to Addgene. .. The following ORF24 fragments: residues 1–201 (ORF24-NTD) (Addgene #138420), residues 1–271 (Addgene #138421), and residues 1–133 (Addgene #138422) were PCR amplified from pcDNA4/TO-ORF24-3xFLAG [ ] (Addgene #138423) with primers to introduce BamHI and NotI sites and cloned into pcDNA4/TO-3xFLAG (C-terminal tag) using T4 DNA ligase (New England Biolabs). pcDNA4/TO-ORF24 202-752-3xFLAG (Addgene #138424) was generated by inverse site-directed mutagenesis with Phusion DNA polymerase (New England Biolabs) using pcDNA4/TO-ORF24-3xFLAG as a PCR template. .. PCR products from inverse PCR were DpnI treated, then ligated using T4 PNK and T4 DNA ligase (New England Biolabs).
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