Table I.
Artificial RNA Standards and Primers for Quantitative RT-PCR
| mRNA | Standard sequence | Upper primer | Lower primer | Buffer | Reference | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| Albumin | 391-584; NcoI (215) | 391-407 | 568-584 | K | Minghetti et al., 1985 | |||||
| Transferrin | 750 bp of 3′ end in vector | 5′-TCA ACC TCA | 5′-GCA GCG AAG | A | Ruppert et al., 1990 | |||||
| pmTF-2; NcoI | CGA CTC CT-3′ | ACT ACA CC-3′ | ||||||||
| Keratin 8 | 1299-3′-end; d1317 1348 | 1299-1316 | 1439-1456 | F | Morita et al., 1988 | |||||
| Keratin 18 | 904-3′-end; d921-952 | 904-920 | 1023-1040 | F | Singer et al., 1986 | |||||
| HNF-1 | 1122-1997; AocI (1715) | 1494-1511 | 1765-1782 | J | Kuo et al., 1990 | |||||
| β-Globin | pGEM3Zf (1357-1550) | 38390-38407 | 39436-39453 | N | Shehee et al., 1989 | |||||
| ε-Globin | pGEM3Zf (1357-1550) | 9625-9642 | 10482-10499 | N | Shehee et al., 1989 | |||||
| Erythropoietin | 1239-3201; AccI (2365) | 1303-1320 | 3149-3166 | N | Shoemaker et al., 1986 | |||||
| c-jun | 1430-1492/811 bp neo/2205-2304 | 1430-1447 | 2287-2304 | A | Ryder and Nathans, 1988 | |||||
| junB | 1081-1483; d1249-1332 | 1081-1098 | 1466-1483 | J | Ryder et al., 1988 | |||||
| junD | pGEM3Zf (1132-1275) | 519-535 | 732-748 | N | Ryder et al., 1989 | |||||
| c-fos | 2969-3574; AccI (3220) | 2969-2992 | 3551-3574 | J | van Beveren et al., 1983 | |||||
| fosB | 6379-6830; d6418-6592 | 6397-6414 | 6813-6830 | J | Lazo et al., 1992 | |||||
| c-myc | 969-1407; d1003-1186 | 969-986 | 1390-1407 | J | Stanton et al., 1984 | |||||
| Cyclin D2 | 55-520; d228-293 | 55-76 | 502-520 | F | Matsushime et al., 1991 | |||||
| Hepatocyte growth factor | 1186-1748; d1384-1595 | 1186-1203 | 1731-1748 | B | Liu et al., 1993 | |||||
| Glyceraldehyde-3-phosphate | 326-1152; CelII (510) | 428-445 | 787-804 | L | Sabath et al., 1990 | |||||
| dehydrogenase |
Bp numbering corresponds to the published cDNA or genomic DNA sequences. Restriction enzymes used for generation of standards with a mutated restriction site, the positions of the deletions in deletion standards (d) and the positions of the heterologous sequences on plasmid pGEM3z, amplified for heterologous standards, are indicated. In the case of the heterologous standard for c-jun, a part of the c-jun sequence between bp 1492 and 2205 is replaced by the neomycin resistance gene sequence of transposon TN5 (Hilberg et al., 1993). The indicated buffers correspond to the various PCR buffers of the PCR optimizer kit (Invitrogen).