Systematic approach to restriction mapping?

Bob Tacon bob_tcc at yahoo.com
Fri Oct 29 21:13:15 EST 1999


Hello!

Is there a systematic approach to restriction mapping? Especially when
you are working with several enzymes it is getting difficult to find the
possible combinations and determine the order of the restriction sites.
All literature I found only uses simple digestions with not more than
two enzymes.

Example: A plasmid was cut with four different enzymes. Double and
triple digestions were also performed. The results are:

Cut with EcoRI                               10
Cut with HindIII                              5 , 3 , 2
Cut with with HaeIII                        6.5 , 3.5
Cut with BgI II                                6 , 4
Cut with HindIII + BgI II                 4 , 3, 2 , 1
Cut with HindIII + HaeIII                5, 2 , and 2 moles of 1.5
Cut with HaeIII + BgI II                  4 , 3.5 , 2.5
Cut with HindIII + HaeIII + BgI II   4, 2 , 2 moles of 1.5 , 1

How would you best approach such a restriction mapping problem?


Bob




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