How many nucleotides are required beyond restriction site by restriction enzymes like NdeI and BamHI for restriction?

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Pierre Béguin

Usually 3 nucleotides is OK for most REs. You can check this on the web site of the suppliers (New England Biolabs, ThermoFisher)

Manuele Martinelli

Dear Muhammad

you can find the flanking sequences suggested from NEB for each enzime at

https://international.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en&rev=c2f94e1cdcd549c5bf8fdb59f7b63f67&hash=4FBF94A3878D7C5E79BC1DBBADD5CCDF70803D06

Ciao

Manuele

Muhammad Shahid Nadeem

Manuele Martinelli, many thanks for sharing the useful page. We have added 8 to 11 bp at the 5' end of our gene sequence and initially it has given better results.