Sequence amplified | Primer sequence | Annealing temp | MgCl2(mM) | No of cycles | Amplicon (bp) | |
Enterovirus 5′ UTR | First round PCR | 50°C | 2.0 | 30 | 228 | |
Nucleotide positions 417–645 | ||||||
5′-GGTGYGAAGAGYCTAYTGAG | ||||||
5′-CACYGGATGGCYATTCCA | ||||||
Nested reaction | 52°C | 1.8 | 25 | 146 | ||
Nucleotide positions 456–602 | ||||||
5′-CCCCTGAATGCGGCTAAT | ||||||
5′-ATTGTCACCATAAGCAGCCA | ||||||
HCMV UL83 region | First round PCR | 57°C | 1.5 | 40 | 449 | |
Nucleotide positions 120032–120489 | ||||||
5′-TCTCGCGTATGGCTTG | ||||||
5′-GTCAGCGTTCGTGTTTCC | ||||||
Nested reaction | 55°C | 1.5 | 30 | 233 | ||
Nucleotide positions 120144–120377 | ||||||
5′-TGAGGTAAAAGCCACATCCAG | ||||||
5′-GACCAGTACGTCAAGGTGTAC | ||||||
Adenovirus hexon region | First round PCR | 60°C | 2.0 | 40 | 300 | |
Nucleotide positions 18858–19158 | ||||||
5′-GCCGCAGTGGTCTTACATGCACATC | ||||||
5′-CAGCACGCCGCGGATGTCAAAGT | ||||||
Nested reaction | 60°C | 2.0 | 30 | 142 | ||
Nucleotide positions 18937–19079 | ||||||
5′-GCCACCGAGACGTACTTCAGCCTG | ||||||
5′-TTGTACGAGTACGCGGTATCCTCGCGGTC | ||||||
Myoglobin exons 1 and 3 | Amplification from DNA | 55°C | 1.5 | 35 | 439 | |
5′-AACATGACAGGTCCTCTTGG | ||||||
5′-CATGGCGCAGTCTGAA | ||||||
Amplification from cDNA | 55°C | 1.5 | 40 | 299 | ||
5′TTGGCAGGAAGTCCTCATCA | ||||||
5′-AGCTCCAGGGCCTTGTTCAT |
bp, base pair; HCMV UL83, human cytomegalovirus unique long; PCR, polymerase chain reaction.