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Conventional and methylation-sensitive gel-blotting analyses followed similar protocols, the only difference lied in using different restriction enzymes for DNA digestion.
However, there was no significant difference in the one-dimensional scatter plots for different restriction enzyme digestion (Fig. 4a).
However, this 2 bp difference did change a HinfI restriction site resulting in a different restriction pattern.
In order to explore the limitation of the method we also computed all possible restricted fingerprints of MCP-2 and Cystatin C containing only 5 different restriction enzymes.
Vectors have target sites for many different restriction enzymes, but the most convenient ones are those that occur only once in the vector molecule.
This suggests a different restriction, on top of the "as presently to be done" restriction.
Firstly, the different restriction of derivative we give is necessary to guarantee the existence of the second-order derivative.
Species identification was performed by digesting mini-exon PCR products with one or two different restriction enzymes.
The homologous ends of the up- and downstream fragments overlapped each other and were flanked by two different restriction sites (KpnI and SpeI).
Two different size markers, 1 kb (Promega, USA) and 100 bp (Promega, USA) DNA ladders were used to analyze different restriction fragments.
Zeschnigk et al. applied a similar approach using 454-sequencing with a combination of 4 different restriction enzymes [26].
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