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very hard 2 step digestion question??

very hard 2 step digestion question?? - Protocols and Methods Forum

very hard 2 step digestion question?? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 02-10-2009, 02:49 AM
Pipette Filler
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Default very hard 2 step digestion question??



this is a question from one of my mgenetics labs im having trouble with

experimental constraints demand that a plasmid be constructed in two digestion steps whose order cannot be reveresed. in step 1, a BamHI fragment is inserted into a BamHI site of plasmid pAMP,

in step 2, an EcoRI fragment must be cloned into an EcorRI site within BamHI insert. Unfortuently the pAmp ''backbone" also contains an EcoRI site, which is not the intended cloning site for the EcoRI fragment in step 2

a) draw a diagram of this cloning experiment

b) explain how EcorRI methylase could be used to solve this experimental proble

any insight into the problem is appreciated
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