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Figure 2 | BMC Genomics

Figure 2

From: The construction and use of bacterial DNA microarrays based on an optimized two-stage PCR strategy

Figure 2

Design and use of chimeric bipartite oligonucleotides for PCR amplification of each open reading frame of the genome from Synechocystis PCC6803. Bipartite oligos (a and b) were designed to include: 1) a gene specific sequence at their 3' end of variable length to maintain a constant tm of 58–62°C; and 2) universal sequence at their 5' to introduce a common sequence to all primary products. These primary products were then re-amplified by universal oligonucleotides (c and d) introducing more universal sequence. The re-amplification of products using universal oligonucleotides allows amplification of all products under optimized conditions, thus improving the success rate and product yields. The final products of the second amplification are analyzed for size, yield, and lack of multiplicity on agarose gels, then purified, resuspended in printing buffer, and printed on modified surface glass slides. This strategy minimizes the cost and effort to replicate the PCR-generated DNA gene fragment library and facilitates several downstream processes beyond the primary objective of producing DNA microarrays.

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