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Nebuilder hifi cloning master mix

Manufactured by New England Biolabs
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NEBuilder HiFi Cloning Master Mix is a reagent designed for seamless DNA assembly. It enables efficient and accurate DNA fragment joining without the need for restriction enzymes or specialized equipment.

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Lab products found in correlation

Pcag egxxfp, by Addgene (2 mentions) Pcdna3.1 vector, by Thermo Fisher Scientific (2 mentions) Zymogen kits, by Genesee Scientific (2 mentions) Pdonr223 tie1 plasmid, by Addgene (2 mentions) Plenti cmv blast vector, by Addgene (2 mentions) Pentr4 vector, by Addgene (2 mentions) Gateway lr clonase kit, by Thermo Fisher Scientific (2 mentions) Px459, by Addgene (2 mentions) Q5 site directed mutagenesis kit, by New England Biolabs (2 mentions)

2 protocols using nebuilder hifi cloning master mix

1

Recombinant Tie1 Extracellular Domain Expression

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All miniprep, maxiprep, and gel extraction steps were conducted using Zymogen kits (Genesee Scientific). Cloning was performed using the NEBuilder HiFi Cloning Master Mix (New England Biolabs) unless otherwise noted. Human Tie1 cDNA was obtained from pDONR223-TIE1 plasmid (23946, Addgene). The Tie1 extracellular domain excluding the native secretion signal (A21 to Q759) was cloned into a pcDNA3.1 vector (ThermoFisher Scientific) containing an N-terminal Ig κ-leader sequence and C-terminal human IgG1 Fc and 6xHis tags. Truncation (Tie1-N: A21 to S431) and site mutation (Tie1-2A, Tie1-6A) constructs were generated using the Q5 site-directed mutagenesis kit (New England Biolabs). The transmembrane HTP construct was produced as previously described.35 (link) To obtain the lentiviral construct, the HTP sequence was first cloned into the pENTR4 vector (17424, Addgene) and then transferred into the pLenti-CMV-Blast vector (17451, Addgene) using the Gateway LR Clonase kit (ThermoFisher Scientific). The vectors pCAG-EGxxFP (50716, Addgene) and pX459 (62988, Addgene) were used to test gRNA sequences.
Griffin M.E., Sorum A.W., Miller G.M., Goddard WA I.I.I, & Hsieh-Wilson L.C. (2020). Sulfated glycans engage the Ang/Tie pathway to regulate vascular development. Nature chemical biology, 17(2), 178-186.
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2

Recombinant Tie1 Extracellular Domain Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
All miniprep, maxiprep, and gel extraction steps were conducted using Zymogen kits (Genesee Scientific). Cloning was performed using the NEBuilder HiFi Cloning Master Mix (New England Biolabs) unless otherwise noted. Human Tie1 cDNA was obtained from pDONR223-TIE1 plasmid (23946, Addgene). The Tie1 extracellular domain excluding the native secretion signal (A21 to Q759) was cloned into a pcDNA3.1 vector (ThermoFisher Scientific) containing an N-terminal Ig κ-leader sequence and C-terminal human IgG1 Fc and 6xHis tags. Truncation (Tie1-N: A21 to S431) and site mutation (Tie1-2A, Tie1-6A) constructs were generated using the Q5 site-directed mutagenesis kit (New England Biolabs). The transmembrane HTP construct was produced as previously described.35 (link) To obtain the lentiviral construct, the HTP sequence was first cloned into the pENTR4 vector (17424, Addgene) and then transferred into the pLenti-CMV-Blast vector (17451, Addgene) using the Gateway LR Clonase kit (ThermoFisher Scientific). The vectors pCAG-EGxxFP (50716, Addgene) and pX459 (62988, Addgene) were used to test gRNA sequences.
Griffin M.E., Sorum A.W., Miller G.M., Goddard WA I.I.I, & Hsieh-Wilson L.C. (2020). Sulfated glycans engage the Ang/Tie pathway to regulate vascular development. Nature chemical biology, 17(2), 178-186.
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