T7 or SP6 - (Dec/16/2008 )

Hi Guys,

May be this is very simple question for you but i am new to moleular biology so trying to clear my concepts.

Can anyone tell me that on what bases do we select the restriction enzyme to digest the plasmid DNA?
Second can we use any restriction enzyme to digest plasmids?
Third how we will determine that should we use T7, SP6 or T3 mMessage Kit to make cRNA?

Hope someone on the forum will help me out with these basic questions.

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Hi,
Selection of restriction enzymes: There are several points to consider when choosing the enzymes
e.g.:
-> You need unique digestion sites for cloning. It doesn't make sense if there are multiple sites in your construct or the vector into which you'd like to clone.
-> Do you have the enzymes already in your freezer? Are they relativley common and easy/cheap available?
-> Do you want to clone something in frame?

Most enzymes are usable for plasmids. For this purpose they were basically provided. Which you can use:
Take the plasmid map and search for one appropriate (see above)

To this message Kit I can't tell you anything. But I guess which Promotor is the right one, the plasmid map will tell you.

I hope this helps a little
grettings
Jazim

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