Ask help for SIRT6 promoter primers design

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4.3 years ago

xxyzoe0828 • 0

Hi, I need to clone the promoter region of SIRT6 to conduct the following luciferase. Could anyone be able to help me about designing the primers for SIRT6 promoter region. As I need to insert this product into the plasmid pGL3, is there anything I need to consider about?

luciferase • 807 views

ADD COMMENT • link 4.3 years ago by xxyzoe0828 • 0

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is there anything I need to consider about?

is it related to bioinformatics ? otherwise, ask https://biology.stackexchange.com/

ADD REPLY • link 4.3 years ago by Pierre Lindenbaum 161k

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thanks for reminding me!

ADD REPLY • link 4.3 years ago by xxyzoe0828 • 0

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Not really, other than throwing on sequences for the restriction enzymes to be used for cloning on the ends of your primers with a few extra bases to aid digestion.

ADD REPLY • link 4.3 years ago by jared.andrews07 ★ 16k

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I strongly recommend restriction-free cloning like FusionHD from Takara or HiFi Assembly from NEB. This works via homology overhangs (15-20nt overhangs) and does not require any digestion of the cloned fragment.