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Hifi 2x assembly master mix

Manufactured by New England Biolabs
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HiFi 2X Assembly Master Mix is a high-fidelity DNA assembly reagent that enables efficient and accurate joining of DNA fragments. It is designed for a variety of applications, including cloning, gene synthesis, and construction of complex genetic circuits.

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Lab products found in correlation

Pcep4, by Thermo Fisher Scientific (2 mentions) Tet inducible promoter, by Takara Bio (2 mentions)

2 protocols using hifi 2x assembly master mix

1

Inducible Genomic Integration of LINE-1

Check if the same lab product or an alternative is used in the 5 most similar protocols
Plasmids used in this study are listed in Supplementary Table 5. The
mammalian expression vector pCEP4 (Invitrogen) was modified to possess a
2nd or 3rd Generation Tet-inducible promoter
(ClonTech) by Gibson assembly. LINE-1 sequences were inserted into the
vector backbone by Gibson assembly with PCR amplicons of endogenous LINE-1
sequence (LINE-1 RP) or ORFeus codon-optimized sequence83 . Control pCEP4 vectors encoded
either eGFP or lacked expression inserts. LINE-1 point mutant constructs
were also created by amplification and Gibson assembly. For sleeping-beauty
integrated LINE-1, ORFeus codon-optimized LINE-1 was cloned into the donor
vector pSBtet-RN or pSBtet-GN84 by Gibson assembly. Briefly, pSBtet-RN or GN was
digested with SfiI and DraIII, gel purified and assembled with PCR-amplified
LINE-1 (primers SB-ORFeus-5 and SB-ORFeus-3 in Supplementary Table 6) using
the HiFi 2X Assembly Master Mix (NEB, Ipswich, MA).
Ardeljan D., Steranka J.P., Liu C., Li Z., Taylor M.S., Payer L.M., Gorbounov M., Sarnecki J.S., Deshpande V., Hruban R.H., Boeke J.D., Fenyö D., Wu P.H., Smogorzewska A., Holland A.J, & Burns K.H. (2020). Cell fitness screens reveal a conflict between LINE-1 retrotransposition and DNA replication. Nature structural & molecular biology, 27(2), 168-178.
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2

Inducible Genomic Integration of LINE-1

Check if the same lab product or an alternative is used in the 5 most similar protocols
Plasmids used in this study are listed in Supplementary Table 5. The
mammalian expression vector pCEP4 (Invitrogen) was modified to possess a
2nd or 3rd Generation Tet-inducible promoter
(ClonTech) by Gibson assembly. LINE-1 sequences were inserted into the
vector backbone by Gibson assembly with PCR amplicons of endogenous LINE-1
sequence (LINE-1 RP) or ORFeus codon-optimized sequence83 . Control pCEP4 vectors encoded
either eGFP or lacked expression inserts. LINE-1 point mutant constructs
were also created by amplification and Gibson assembly. For sleeping-beauty
integrated LINE-1, ORFeus codon-optimized LINE-1 was cloned into the donor
vector pSBtet-RN or pSBtet-GN84 by Gibson assembly. Briefly, pSBtet-RN or GN was
digested with SfiI and DraIII, gel purified and assembled with PCR-amplified
LINE-1 (primers SB-ORFeus-5 and SB-ORFeus-3 in Supplementary Table 6) using
the HiFi 2X Assembly Master Mix (NEB, Ipswich, MA).
Ardeljan D., Steranka J.P., Liu C., Li Z., Taylor M.S., Payer L.M., Gorbounov M., Sarnecki J.S., Deshpande V., Hruban R.H., Boeke J.D., Fenyö D., Wu P.H., Smogorzewska A., Holland A.J, & Burns K.H. (2020). Cell fitness screens reveal a conflict between LINE-1 retrotransposition and DNA replication. Nature structural & molecular biology, 27(2), 168-178.
+ Open protocol
+ Expand

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