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. Author manuscript; available in PMC: 2010 Oct 20.
Published in final edited form as: Methods Enzymol. 2010 Mar 1;470:181–204. doi: 10.1016/S0076-6879(10)70008-2

Fig. 5. Testing the Topo-TA cloning efficiency in SB221.

Fig. 5

Unless otherwise stated, all the symbols are as in Fig. 3 (A.) The SB221 vector is made up of the URA3 gene flanked by a fragment containing the TEF promoter plus half of the KanMX gene (Kan), and another fragment containing the other half of the KanMX gene and the TEF terminator (Mx). This 2.7kb fragment can be excised from the 2.5kb backbone by restriction digestion with NotI. (B.) Restriction digest with NotI following a successful Topo-TA cloning reaction is indicated by the 2.5kb backbone and a band shift of the 2.7kb fragment, in accordance with the insert size.