Primer pairs and cycling conditions for Lightcycler real-time PCR
| PCR primer pair . | Origin . | Sequence . | Denaturing temperature . | Annealing temperature (°C)/time . | Extension temperature (°C)/time . | Acquisition temperature (°C)/time . |
|---|---|---|---|---|---|---|
| Antipodean | Stennard et al., 1999 | U: 5′-TGG ATT AGT TTA GGA ACA -3′ | 95 | 55/5 | 72/16 | 83/3 |
| D: 5′-CGG ATC TTA CAC TGA GGA -3′ | ||||||
| Antivin | New | U: 5′-GCC TCA AAG GGA TGT GGA G-3′ | 95 | 55/5 | 72/11 | 81/3 |
| D: 5′-TGT GGC TGA GTC AAC GAG AG-3′ | ||||||
| Bix1 | Xanthos et. al., 2001 | U: 5′-AGA GAC TCC CAG TTC ATC TGA-3′ | 95 | 60/5 | 72/8 | 83/3 |
| D: 5′-GGT AGG TGG GAA GTT GCT AAT-3′ | ||||||
| Bix3 | Xanthos et. al., 2001 | U: 5′-TCT CGC ATT CAG GTT TGG TTC C-3′ | 95 | 60/5 | 72/11 | 83/3 |
| D: 5′-ATC TCC TTG TTA GGG ATC ATA C-3′ | ||||||
| Bix4 | Xanthos et. al., 2001 | U: 5′-AGA TGC TAC AGG CTG GAG CAA-3′ | 95 | 60/5 | 72/11 | 84/3 |
| D: 5′-GTG TGT AAG GGG TGA GTC ATA-3′ | ||||||
| Cerberus | Heasman et al., 2000 | U: 5-GCT TGC AAA ACC TTG CCC TT-3′ | 95 | 60/5 | 72/20 | 81/3 |
| D: 5-CTG ATG GAA CAG AGA TCT TG-3 | ||||||
| Eomesodermin | New | U: 5′-TGG TCC TCA AGG TCA AGT CC-3′ | 95 | 56/6 | 72/10 | 83/3 |
| D: 5′-GGG GAG TTT TCA TTG CTT GA-3′ | ||||||
| Endodermin | Sasai et al., 1996 | U: 5′-TAT TCT GAC TCC TGA AGG TG-3′ | 95 | 55/5 | 72/8 | 81/3 |
| D: 5′-GAG AAC TGC CCA TGT GCC TCC-3′ | ||||||
| Epidermal keratin | XMMR* | U: 5′-CAC CAG AAC ACA GAG TAC -3′ | 95 | 55/5 | 72/9 | 81/3 |
| D: 5′-CAA CCT TCC CAT CAA CCA -3′ | ||||||
| eFGF | New | U: 5′-TAT GAA TGC AAA GGG GAA GC-3′ | 95 | 55/5 | 72/10 | 83/3 |
| D: 5′-ATA TCC GTG GCA AGA AAT GG-3′ | ||||||
| Fgf3 | Kofron et al., 1999 | U: 5′-GTC ATT TGT TTC CAG ACT TC-3′ | 95 | 55/5 | 72/12 | 85/3 |
| D: 5′-TAT CTG TAG GTG GTA CTT AG-3′ | ||||||
| Fgf8 | Kofron et al., 1999 | U: 5′-CTG GTG ACC GAC CAA CTA AG-3′ | 95 | 55/5 | 72/14 | 86/3 |
| D: 5′-ACC AGC CTT CGT ACT TGA CA-3′ | ||||||
| Gata2 | New | U: 5′-CTA AAC AGA GGA GCA AGA GC-3′ | 95 | 55/5 | 72/15 | 86/3 |
| D: 5′-CCT AAG TTC CTC AAA AC-3′ | ||||||
| Gata5 | Xanthos et. al., 2001 | U: 5′-ACC TTC AGA GCT GCG ACA CT-3′ | 95 | 60/5 | 72/20 | 86/3 |
| D: 5′-CCA ACC GGG AGC CCC GAT A-3′ | ||||||
| ODC | Heasman et al., 2000 | U: 5′-GCC ATT GTG AAG ACT CTC TCC ATT C-3′ | 95 | 55/5 | 72/12 | 83/3 |
| D: 5′-TTC GGG TGA TTC CTT GCC AC-3′ | ||||||
| Sprouty2 | New | U: 5′-TGC TCA GCC CAG GAA GTA GT-3′ | 95 | 55/5 | 72/10 | 83/3 |
| D: 5′-ACC ACA AAC AGG GCA AGA AC-3′ | ||||||
| Xbra | Sun et al., 1999 | U: 5′-TTC TGA AGG TGA GCA TGT CG-3′ | 95 | 55/5 | 72/8 | 75/3 |
| D: 5′-GTT TGA CTT TGC TAA AAG AGA CAG G-3′ | ||||||
| Xlim1 | New | U: 5′-CCCTGGCAGCAACTATGACT-3′ | 95 | 55/5 | 72/11 | 85/3 |
| D: 5′-GGTTGCCATAACCTCCATTG-3′ | ||||||
| Xlim5 | Houston et. al., 2003 | U: 5′ - CCA ACA GAC AGG CCC AAC-3′ | 95 | 60/5 | 72/8 | 84/3 |
| D: 5′-GTG GCT CCG GTG CTA CAG - 3′ | ||||||
| Xnr1 | Kofron et al., 1999 | U: 5′-TGG CCA GAT AGA GTA GAG-3′ | 95 | 55/5 | 72/12 | 81/3 |
| D: 5′-TCC AAC GGT TCT CAC TTT-3′ | ||||||
| Xnr5 | New | U: 5′-ATG AGG CCT CTG TCA ATG CT-3′ | 95 | 55/5 | 72/10 | 80/3 |
| D: 5′-GCC CTG AAT GTC TTG CAT CT-3′ | ||||||
| Xnot1 | New | U: 5′-CTGCATTTGGCCACCACCTGGC-3′ | 95 | 55/5 | 72/15 | 86/3 |
| D: 5′-GATGAGCCACACGGGTGGGTA-3′ | ||||||
| Xsox17 | Xanthos et al., 2001 | U: 5′-GCA AGA TGC TTG GCA AGT CG-3′ | 95 | 58/5 | 72/8 | 85/3 |
| D: 5′-GCT GAA GTT CTC TAG ACA CA-3′ |
| PCR primer pair . | Origin . | Sequence . | Denaturing temperature . | Annealing temperature (°C)/time . | Extension temperature (°C)/time . | Acquisition temperature (°C)/time . |
|---|---|---|---|---|---|---|
| Antipodean | Stennard et al., 1999 | U: 5′-TGG ATT AGT TTA GGA ACA -3′ | 95 | 55/5 | 72/16 | 83/3 |
| D: 5′-CGG ATC TTA CAC TGA GGA -3′ | ||||||
| Antivin | New | U: 5′-GCC TCA AAG GGA TGT GGA G-3′ | 95 | 55/5 | 72/11 | 81/3 |
| D: 5′-TGT GGC TGA GTC AAC GAG AG-3′ | ||||||
| Bix1 | Xanthos et. al., 2001 | U: 5′-AGA GAC TCC CAG TTC ATC TGA-3′ | 95 | 60/5 | 72/8 | 83/3 |
| D: 5′-GGT AGG TGG GAA GTT GCT AAT-3′ | ||||||
| Bix3 | Xanthos et. al., 2001 | U: 5′-TCT CGC ATT CAG GTT TGG TTC C-3′ | 95 | 60/5 | 72/11 | 83/3 |
| D: 5′-ATC TCC TTG TTA GGG ATC ATA C-3′ | ||||||
| Bix4 | Xanthos et. al., 2001 | U: 5′-AGA TGC TAC AGG CTG GAG CAA-3′ | 95 | 60/5 | 72/11 | 84/3 |
| D: 5′-GTG TGT AAG GGG TGA GTC ATA-3′ | ||||||
| Cerberus | Heasman et al., 2000 | U: 5-GCT TGC AAA ACC TTG CCC TT-3′ | 95 | 60/5 | 72/20 | 81/3 |
| D: 5-CTG ATG GAA CAG AGA TCT TG-3 | ||||||
| Eomesodermin | New | U: 5′-TGG TCC TCA AGG TCA AGT CC-3′ | 95 | 56/6 | 72/10 | 83/3 |
| D: 5′-GGG GAG TTT TCA TTG CTT GA-3′ | ||||||
| Endodermin | Sasai et al., 1996 | U: 5′-TAT TCT GAC TCC TGA AGG TG-3′ | 95 | 55/5 | 72/8 | 81/3 |
| D: 5′-GAG AAC TGC CCA TGT GCC TCC-3′ | ||||||
| Epidermal keratin | XMMR* | U: 5′-CAC CAG AAC ACA GAG TAC -3′ | 95 | 55/5 | 72/9 | 81/3 |
| D: 5′-CAA CCT TCC CAT CAA CCA -3′ | ||||||
| eFGF | New | U: 5′-TAT GAA TGC AAA GGG GAA GC-3′ | 95 | 55/5 | 72/10 | 83/3 |
| D: 5′-ATA TCC GTG GCA AGA AAT GG-3′ | ||||||
| Fgf3 | Kofron et al., 1999 | U: 5′-GTC ATT TGT TTC CAG ACT TC-3′ | 95 | 55/5 | 72/12 | 85/3 |
| D: 5′-TAT CTG TAG GTG GTA CTT AG-3′ | ||||||
| Fgf8 | Kofron et al., 1999 | U: 5′-CTG GTG ACC GAC CAA CTA AG-3′ | 95 | 55/5 | 72/14 | 86/3 |
| D: 5′-ACC AGC CTT CGT ACT TGA CA-3′ | ||||||
| Gata2 | New | U: 5′-CTA AAC AGA GGA GCA AGA GC-3′ | 95 | 55/5 | 72/15 | 86/3 |
| D: 5′-CCT AAG TTC CTC AAA AC-3′ | ||||||
| Gata5 | Xanthos et. al., 2001 | U: 5′-ACC TTC AGA GCT GCG ACA CT-3′ | 95 | 60/5 | 72/20 | 86/3 |
| D: 5′-CCA ACC GGG AGC CCC GAT A-3′ | ||||||
| ODC | Heasman et al., 2000 | U: 5′-GCC ATT GTG AAG ACT CTC TCC ATT C-3′ | 95 | 55/5 | 72/12 | 83/3 |
| D: 5′-TTC GGG TGA TTC CTT GCC AC-3′ | ||||||
| Sprouty2 | New | U: 5′-TGC TCA GCC CAG GAA GTA GT-3′ | 95 | 55/5 | 72/10 | 83/3 |
| D: 5′-ACC ACA AAC AGG GCA AGA AC-3′ | ||||||
| Xbra | Sun et al., 1999 | U: 5′-TTC TGA AGG TGA GCA TGT CG-3′ | 95 | 55/5 | 72/8 | 75/3 |
| D: 5′-GTT TGA CTT TGC TAA AAG AGA CAG G-3′ | ||||||
| Xlim1 | New | U: 5′-CCCTGGCAGCAACTATGACT-3′ | 95 | 55/5 | 72/11 | 85/3 |
| D: 5′-GGTTGCCATAACCTCCATTG-3′ | ||||||
| Xlim5 | Houston et. al., 2003 | U: 5′ - CCA ACA GAC AGG CCC AAC-3′ | 95 | 60/5 | 72/8 | 84/3 |
| D: 5′-GTG GCT CCG GTG CTA CAG - 3′ | ||||||
| Xnr1 | Kofron et al., 1999 | U: 5′-TGG CCA GAT AGA GTA GAG-3′ | 95 | 55/5 | 72/12 | 81/3 |
| D: 5′-TCC AAC GGT TCT CAC TTT-3′ | ||||||
| Xnr5 | New | U: 5′-ATG AGG CCT CTG TCA ATG CT-3′ | 95 | 55/5 | 72/10 | 80/3 |
| D: 5′-GCC CTG AAT GTC TTG CAT CT-3′ | ||||||
| Xnot1 | New | U: 5′-CTGCATTTGGCCACCACCTGGC-3′ | 95 | 55/5 | 72/15 | 86/3 |
| D: 5′-GATGAGCCACACGGGTGGGTA-3′ | ||||||
| Xsox17 | Xanthos et al., 2001 | U: 5′-GCA AGA TGC TTG GCA AGT CG-3′ | 95 | 58/5 | 72/8 | 85/3 |
| D: 5′-GCT GAA GTT CTC TAG ACA CA-3′ |
Xenopus Molecular Marker Resource(http://www.xenbase.org/xmmr/Marker_pages/primers.html).