Multiplex PCR examinations of RCA amplified whole genome. ( a ) Control, multiplex PCR amplifications for the 12 randomly selected human genes using human genomic DNA as the template. Amount of the template DNA for the PCR is indicated. Multiplex PCR amplifications of the 12 human genes using RCA products as the template in the absence ( b ) or presence ( c ) of Tth SSB-255 protein. Amount of the template DNA for RCA is serially diluted as indicated. Throughout (a–c), all samples were amplified with primers at the same primer pair concentrations (0.1 µM per pair). Aliquots of 5 µl volume were electrophoresed through 12.5% acrylamide gel in Tris–borate/EDTA buffer, and stained with SYBR Green (SYBR Green I, Novagen). The signals were detected using a Fluoro Imager (Fluoro Imager 595, Molecular Dynamics). Product sizes (from 57 to 360 bp) are indicated on the right or left side of each panel. The oligonucleotide sequences used for the primers are as follows. Primer sets 1 (chromosome 11), 5′-GGGCA GAGCC ATCTA TTGCT TACA-3′, 5′-GGTTG CTAGT GAACA CAGTT GTGTC A-3′; Primer sets 2 (chromosome 16), 5′-GCACT CTTCT GGTCC CCACA GA, 5′-TTGGT CTTGT CGGCA GGAGA CA-3′; Primer sets 3 (chromosome 8), 5′-GTCCT TCCCC CGCTG GAAAC-3′, 5′-GCAGC AGAGA TCATC GCGCC-3′; Primer sets 4 (chromosome 7), 5′-CACAG ATTTC CAAGG ATGCG CTG, 5′-CGTGC TCTGT TCCAG ACTTG-3′; Primer sets 5 (chromosome 10), 5′-CGTCT GGCGA TTGCT CCAAA TG-3′, 5′-GGGCA GTTGT GATCC ATGAG AA-3′; Primer sets 6 (chromosome 17), 5′-GCCTC TGATT CCTCA CTGAT TGCTC T, 5′-TGTCA ACCAC CCTTA ACCCC TCC-3′; Primer sets 7 (chromosome 20), 5′-TTGGA GGGGT GGGTG AGTCA AG, 5′-GGAGG GGTGG GGGTT AATGG TTA-3′; Primer sets 8 (chromosome 13), 5′-GGAAC AAGAC ACGGC TGGGT T-3′, 5′-AGCAA GGCAG GGCAG GCAAG T-3′; Primer sets 9 (chromosome 3), 5′-CGGTC CCATT CTCAG GGAAT CT-3′, 5′-GCCCA GAGGA AGAAG AAGGA AA-3′; Primer sets 10 (chromosome 1), 5′-GCCCC CACCC AGGTT GGTTT CTA-3′, 5′-ATGCC TTCAT CTGGC TCAGT GAA-3′; Primer sets 11 (chromosome 6), 5′-GCTCA GCATG GTGGT GGCAT AA-3′, 5′-CCTCA TACCT TCCCC CCCAT TT-3′; Primer sets 12 (chromosome 22), 5′-GACTA CTCTA GCGAC TGTCC ATCTC-3′, 5′-GACAG CCACC AGATC CAATC-3′.
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