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Figure 2 | BMC Biochemistry

Figure 2

From: Bifunctional TaqII restriction endonuclease: redefining the prototype DNA recognition site and establishing the Fidelity Index for partial cleaving

Figure 2

TaqIl cleavage patterns of DNA substrates, containing 5'-CACCCA-3' and 5'-GACCGA-3' recognition sites. (A) Digestion of the PCR DNA fragment with a single 5'-GACCGA-3' or 5'-CACCCA-3' site. Digestion of 0.3 μg (= 1.2-pmol recognition sites) PCR(GACCGA) substrate (lanes 1-3) and PCR(CACCCA) substrate (lanes 4-6) with 12 pmol TaqII, electrophoresed on a 15% polyacrylamide/TBE gel. Lane M, Sigma PCR 20-bp Low Ladder (selected bands marked); lane 1, undigested PCR(GACCGA) fragment; lane 2, digestion in the optimized buffer; lane 3, digestion in the buffer according to Barker et al. [18]; lane 4, undigested PCR(CACCCA) fragment; lane 5, digestion in the optimized buffer; lane 6, digestion in the buffer according to Barker et al. [18]. (B) TaqII digestion of the PCR DNA fragment with two convergent sites 5'-CACCCA-3'. Digestion of 0.3 μg (= 2.4-pmol recognition sites) PCR(CACCCA/CACCCA) substrate with 24 pmol TaqII. Lane M, Sigma PCR 20-bp Low Ladder (selected bands marked); lane 1, undigested PCR fragment; lane 2, digestion in the optimized buffer.

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BMC Biochemistry

ISSN: 1471-2091

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