How far 2 restriction sites should be? - how many bases in between for efficient digestion. (Jun/07/2006 )
hello again...
so im designing primers for PCR insert to insert it after that in his vector. i have chosen sites of ECoRV and BGLII (have anyone got problems with those?? 
)but they are 12 bases apart... 
....is it enough? im concerned they are too close......what do you think? thankx a lot!
-Kathy-
i guess this will help:
http://www.neb.com/nebecomm/tech_reference...ized_vector.asp
depending which enzyme cuts second, this will be "close to end"
as far as I see, this should be no problem. 12 bases should be no prob anyway with standard nucleases
-Kersten-
EcoRV in combination with BglII worked very well in my restriction digests!
-Jou-
QUOTE (Kersten @ Jun 7 2006, 06:25 AM)
i guess this will help:
http://www.neb.com/nebecomm/tech_reference...ized_vector.asp
depending which enzyme cuts second, this will be "close to end"
as far as I see, this should be no problem. 12 bases should be no prob anyway with standard nucleases
http://www.neb.com/nebecomm/tech_reference...ized_vector.asp
depending which enzyme cuts second, this will be "close to end"
as far as I see, this should be no problem. 12 bases should be no prob anyway with standard nucleases
Should be no problem. If either one cuts first there should be a 12 base overhang either way. That's more than enough for the subsequent cut.
-vasussci-
thanx a lot everyone! 
-Kathy-