URMC / Microbiology & Immunology / Research / Xenopus Laevis / Primers Available

Primers Available

Conventional PCR primer list for X. laevis house keeping and other genes

Primers name	Primer direction and/or sequence a	Size (bp)	TM (oC)
Adult beta-globin	F: 5’ - GACAGCACATGATCGTCAGC- 3’ F: 5’ - CTTCATGGAAGTGGCGAGCC- 3’	58	351
EF-1α	F: 5’ - CCTGAATCACCCAGGCCAGATTGGTG - 3’ R: 5’ - GAGGGTAGTGTGAGAAGCTCTCCACG - 3’	250	56
GAPDH	F: 5’ - ACCCCTTCATCGACTTGGAC - 3’ R: 5’ - GGAGCCAGACAGTTTGTAGTG - 3’	300	55
Gp96	F: 5’ - GCGTTGGTTTCTATTCTGCCTTC- 3’ F: 5’ - TGTCTCAGTCTTGCTGCTCCACAC- 3’	56	400
Hsp72	F: 5’ - AGACTGATCGGGAGAAAGTTC - 3’ R: 5’ - CTTTTCCACAGGTTCCAAGGTACCC - 3’	58	747
Hsc73	F: 5’ - TGGAGTAGAAAGGAATGTACTGATC - 3’ R: 5’ - AACTTCATTGCATTTCTCCAG - 3’	58	1174
LMPX	F: 5’ - TATCAGTGTCTTCCGCCTCC - 3’ R: 5’ - CAGCCGTCCTCCTTCATGTG - 3’	56-64	300

Conventional PCR primer list for Xenopus laevis immune receptor genes

Primer	Sequence (5’-3’ forward/reverse)	Tm (°C)	Size (bp)
b2-microglobulin consensus	F: 5’ - TGACGGTGAATCCTGGAGAC- 3’ R: 5’ - CGATAGCCGTGACAATGAGC- 3’	57	800
CD4	F: 5’ - TCCATCTCTGACATCCCCTC- 3’ R: 5’ - TCACCAGACACACGTCCATT- 3’	60	895
CD8 a	F: 5’ - AAGCCACCTACGACTACCACCAA- 3’ R: 5’ - CCGTTCTTCTCAGTCTCAGGCAC- 3’	59	247
CD8 β	F: 5’ - TCATCATCTCTTTCTGGGGC- 3’ R: 5’ - AATTCAGTGGGTGCTTCCTG- 3’	60	604
CTX	F: 5’ - GCAgcagcggtaatcggag- 3’ R: 5’ - ctcagcatggtcatggaattg- 3’	58	220
Ig Y	F: 5’ - ATGGAGGAGCCAACTCACCATC- 3’ R: 5’ - AAGGTGGCAGCAAGGTGATTG- 3’	58	720
Ig M	F: 5’ - TCTTCTCTTGTAAGGAACCCG- 3’ R: 5’ - GCAATGCCAAACACCTGG- 3’	58	350
MHC class Ia f strain	F: 5’ - GAGTTCTTTGCCCTGGATACA- 3’ R: 5’ - CACAATAAGTTTCTCCTTCAGG- 3’	61	400
MHC class Ia j strain	F: 5’ - GAGTTCATAGCGCTGGATACA- 3’ R: 5’ - CCTCACTGGGTGTAATCTCAG- 3’	61	400
MHC II a chain	F: 5’ - TCAAAGTCAGTGGTTTGGACG- 3’ R: 5’ - GTGTAAATATCATGTTCATTTG- 3’	61	390
MHC II b chain	F: 5’ - ACGGCACCGACAATGTCAGG- 3’ TTAATGGGTGTCTCCAGCATC- 3’	61	450
TCR a	F: 5’ - TTCAGGGACCTCTGGTTGCTTC- 3’ R: 5’ - ACACATCAGCCTCAATCCATCC- 3’	60
TCR V b 1	F: 5’ - CACCCAGGAGCCAAGATC- 3’	56	700
TCR V b 3	F: 5’ - TGACGGTGAATCCTGGAGAC- 3’	58	700
TCR V b 6	F: 5’ - GTCAGCTGACATTCCTTGTAATC- 3’	58	700
TCR V b 8	F: 5’ - GACCAAAAGTTCTTAGCG- 3’	50	700
TCR b constant	R: 5’ - CGATAGCCGTGACAATGAGC- 3’	50-58	700
TCR g	F: 5’ - AGAAGTTCTCAAGCAGGCAG- 3’ R: 5’ - AAGTCAGCTGAAGAGGATCC- 3’	60	328
XNC 2	F: 5’ - GGTATCAAACCGTATCTATGGTG- 3’ R: 5’ - TCTCTATATACTCCTTACTATCAG- 3’	59	450
XNC 6	F: 5’ - CTTTGCATGATCGTAATG- 3’ R: 5’ - CTGCATATCTAGATGGCTTG- 3’	52	386
XNC 7	F: 5’ - CCGATGTGAGCCTTTGTTTCAG- 3’ R: 5’ - GTGCACTCATTTTCCATGAATAGT- 3’	59	380
XNC 11	F: 5’ - CGATGACTTCTTCACCGACC- 3’ R: 5’ - CCTTGCTCCACTCTGGTATG- 3’	58	350

Conventional PCR primer list for X. laevis cytokine and other genes

Primers name	Primer direction and/or sequence a	Size (bp)	TM (oC)
AID	F: 5’ - TCACGACCCCCATAGGAACTAC - 3’ R: 5’ - TTAGGAGACTTTGCCTCAAGG - 3’	450	58
Arginase I	F: 5’ - CTTACAATGAGTAGTAGCCAAGG - 3’ R: 5’ - GCAGAGAAAATGAGGATTCG - 3’	378	56
IL-1β	F: 5’ - AACAGAAGATGGCCAAGACTC - 3’ R: 5’ - ATGCAACCGATTCAAAGC - 3’	150	60
Type II IFN (gamma)	F: 5’ - CTGAGGAAATACTTTAACTCCATTGACC - 3’ R: 5’ - TTGTAACATCTCCCACCTGTATTGTC - 3’	300	60
MX1	F: 5’ - AGGAGTGGTCAACAGGAGCC- 3’ R: 5’ - TGTTCCGCCGCTGTTCCTCT- 3’	829	68
MX2	F: 5’ - GGAACGCCGCACTTGCAGAA- 3’ R: 5’ - CGATTAATCCTGGCACCTCC- 3’	274	68
TNF-α	F: 5’ - GCTCAAGGATAACTCCATCG - 3’ R: 5’ - AACCAAGTGGCACCTGAATG - 3’	203	59
Rag1	F: 5’ - GCGCCAAGAATCTGTGTCACT- 3’ R: 5’ - GTTCTGTTTCATGGTTGTCTACCA- 3’	350	58
Mycobacterium marinum	F: 5’ - AGAGTTTGATCCTGGCTCAG - 3’ R: 5’ - CACGCTCACAGTTAAGCCGT - 3’	378	56
DM-W (sex determination)	F: 5’ - CCACACCCAGCTCATGTAAAG- 3’ R: 5’ - GGGCAGAGTCACATATACTG- 3’	260	52

Conventional PCR primer list for pathogens of X. laevis

Primers name	Primer direction and/or sequence a Ranavirus FV3	Size (bp)	TM (oC)
vDNA polymerase II (60R)	F: 5’ - ACGAGCCCGACGAAGACTACATAG - 3’ R: 5’ - TGGTGGTCCTCAGCATCCTTTG - 3’	378	56
MCP	F: 5’ - GACTTGGCC ACTTATGAC - 3’ R: 5’ - GTCTCTGGAGAAGAAGAAGAA - 3’	525	52
vCARD (64R)	F: 5’ - AGACCCTCCAGAACCTGATTGAC - 3’ R: 5’ - TCTCCCTTCTTGGTGACGCTGT - 3’	150	59
vβHSD (52L)	F: 5’ - CACTGTCCACGATCACGTCTACC - 3’ R: 5’ - AACTTTACGCCCGTCACCCTCA - 3’	550	60
vICP-18 (18K)	F: 5’ - ATGATCCAAGCCTACCTGTGC- 3’ R: 5’ - AAATGTCCTAATCTATACACC- 3’	450	52
veIF-2α (vIF-2α)	F: 5’ - CTGTAAATGTTTAGACTTGGAGTG- 3’ R: 5’ - ACACAAAGGGGCACAGTCCTCC- 3’
53R	F: 5’ - ATCAGTCACAGGTCTTTAGCATAG- 3’ R: 5’ - TCTGCTTGATCTCGGGACACAATG- 3’	441	56
EGFP	F: 5’ - ACGGCCACAAGTTCAGCGTG - 3’ R: 5’ - GTCCATGCCGAGAGTGATCC - 3’	350	56
M. marinum	F: 5’ - AGAGTTTGATCCTGGCTCAG - 3’ R: 5’ - CACGCTCACAGTTAAGCCGT - 3’	378	56
Chytrid	F: 5’ - AGCCAAGAGATCCGTTGTCAAA- 3’ R: 5’ - CCTTGATATAATACAGTGTGCCATATC- 3’	250	60
CTX	F: 5’ - GCAGCAGCGGTAATCGGAG- 3’ R: 5’ - CTCAGCATGGTCATGGAATTG- 3’	220	58

Conventional Q-PCR primer list for X. laevis genes

Primers	Primer direction and/or sequence
GAPDH	F: 5’ - GACATCAAGGCCGCCATTAAGACT - 3’ R: 5’ - AGATGGAGGAGTGAGTGTCACCAT - 3’
EF-1α	F: 5’ - GCCTTCCCAGGGTTCAACAT - 3’ R: 5’ - TCGGTGGGTCACTCTTTGAAT - 3’
GCSF-R	F: 5’ - ACGTGCCAGCTA AACCTCACAGAT - 3’ R: 5’ - TGA CACAGCCTGGGCGAGAAATAA - 3’
IL-1β	F: 5’ - CATTCCCATGGAGGGCTACA - 3’ R: 5’ - TGACTGCCACTGAGCAGCAT - 3’
IL-10	F: 5’ - TGCTGGATCTTAAGCACACCCTGA - 3’ R: 5’ - TGTACAGGCCTTGTTCACGCATCT - 3’
Type I IFN	F: 5’ - GCTGCTCCTGCTCAGTCTCA- 3’ R: 5’ - GAAAGCCTTCAGGATCTGTGTGT- 3’
Type II IFN (gamma)	F: 5’ - CTGAGGAAATACTTTAACTCCATTGACC - 3’ R: 5’ - TTGTAACATCTCCCACCTGTATTGTC- 3’
Type III IFN (lambda)	F: 5’ - TCCCTCCCAACAGCTCATG- 3’ R: 5’ - CCGACACACTGAGCGGAAA - 3’
IFN lambda receptor	F: 5’ - GGAGCCTGATCCCAATGAATTA- 3’
(IFNLR1)	R: 5’ - TCTCAAAGCGCACACTAAGG- 3’
MCSF-R	F: 5’ - TGTATTCTTTGG ACT TGC CGT ATCTGG - 3’ R: 5’ - TTGTTTAGCTTCAAATTCTGGGTAATA - 3
MX1	F: 5’ - AGCAGTGGTCAACAGGAGCC - 3’ R: 5’ - TGTTCCGCCGCTGTTCCTCT - 3
MX2	F: 5’ - GGAACGCCGCACTTGCAGAA - 3’ R: 5’ - CGATTAATCCTGGCACCTCC - 3
TNF-α	F: 5’ - TGTCAGGCAGGAAAGAAGCA - 3’ R: 5’ - CAGCAGAGCAAAGAGGATGGT- 3’
HSP70	F: 5’ - CTCATCAAGCGCAACACAAC - 3’ R: 5’ - CTCCCTCAAACACCTGGATAAG- 3’
Glucocorticoids (GCs) (CYP11A1)	F: 5’ - GGAGGAGTAGACACGACATCTA - 3’ R: 5’ - GGCAGTAAGATCCTTTCCAGAG- 3’
Thyroid hormone receptor α (THRα)	F: 5’ - GTTCTGGATGGAATTACG - 3’ R: 5’ - CATACATTGGCTGTTCTT- 3’
IL-34	F: 5’ - AGCTCTTCTACGTGTATTCCTTGG - 3’ R: 5’ - TGATAAGCGATTGACCTACCTGGG - 3’
iNOS	F: 5’ - AACCGTAAGCCAAAGAAGGA - 3’ R: 5’ - TGGTTCTGGCAGCCACAGT - 3’
FoxP3 (Forkhead box P3)	F: 5’ - AGTGGTGGGCAGTTGTATG - 3’ R: 5’ - AGGTTCCGCATGTAGAAGATG - 3’
IL-1β (interlukin 1 beta)	F: 5’ - CATTCCCATGGAGGGCTACA - 3’ R: 5’ - TGACTGCCACTGAGCAGCAT - 3’
IL-4/13	F: 5’ - GCAATTAACGCTTCCCTTTCC - 3’ R: 5’ - AGAAGATCGTCCCTGACTGA - 3’
IL-6 (Interlukin 6)	F: 5’ - CTGTGGAGTCCTTACAGTACAAG - 3’ R: 5’ - TGGTTCCTTTGTCATCACTCTC - 3’
IL-10 (Interlukin 10)	F: 5’ - TGCTGGATCTTAAGCACACCCTGA - 3’ R: 5’ - TGTACAGGCCTTGTTCACGCATCT - 3’
IL-12β (Interlukin 12 beta)	F: 5’ - GAAACCTGCTGTCCTTCTCTT - 3’ R: 5’ - TCATGGCAGTTCACCTCATATT - 3’
IL-15 SET 1 (Interlukin 15)	F: 5’ - TCTCAACATCTCACCCACATTC - 3’ R: 5’ - ACTCTTCAAATGCCGCTATAAATTC - 3’
IL-15 SET2	F: 5’ - GAAAGCTAAAGATACAGGAGAACAAAG - 3’ R: 5’ - TGTGTGAATGTGGGTGAGATG - 3’
IL-18 (Interlukin 18)	F: 5’ - CAGCAAGTGTTCTGACAACAAG - 3’ R: 5’ - TTCACTCGCTTGAGACTGTATG - 3’
TGFβ SET 1 (Transforming growth factor beta)	F: 5’ - CCCACAGGCCAAAGATATAGAC - 3’ R: 5’ - CATCAGGTAGGGTTTCGTGTT - 3’
TGFβ SET 2	F: 5’ - CTACCTGTAAGGCTGTGGATATG - 3’ R: 5’ - CTGTCATCTTCTCGCTGTCTAC - 3’
CCR-2 SET 1	F: 5’ - ATTGGGCAGAACTGTGGTAG - 3’ R: 5’ - GGGCGAGTAATCTGAGTCATAA - 3’
CCR-2 SET 2	F: 5’ - GTCCCATGATGTCAGAGAGTAAC - 3’ R: 5’ - GTGTGGGCAGAGGCAATA - 3’
Pathogens
FV3 vDNA Polymerase II	F: 5’ - AACCCTCGCCTGGCATCTA - 3’
(60R)	R: 5’ - TGGTGGTCCTCAGCATCCT - 3’
FV3-PKR	F: 5’ - GCTCACCGGCGGGATTA - 3’ R: 5’ - TTCAACTTTATTCATGCGTGCTATC - 3’
M. marinum	F: 5’ - AGAGTTTGATCCTGGCTCAG- 3’ R: 5’ - CACTCGAGTATCTCCGAAGA - 3’
FV3-MCP-90R (Major Capsid Protein)	F: 5’ - GTCCTTTAACACGGCATACCT- 3’ R: 5’ - ATCGCTGGTGTTGCCTATC - 3’
FV3-94L (Clostridium tetani)	F: 5’ - CGGGACCTCTATGAGCTTTATG- 3’ R: 5’ - CTACGAGTACGTCTCTGTGTTC - 3’
Immediate-early protein ICP-18K (82R) – immediate early	F: 5’ - TGTGTCTGGAGAACCCTACA - 3’ R: 5’ - TGGTGATCTTGACTTGAAACTC - 3’
93L – late unknown function	F: 5’ - GCCAAGACCGAGGACAC - 3’ R: 5’ - GGACAAACCAACAGTAGAAACG - 3’
DNA Repair protein RAD2 (95R) –delayed early	F: 5’ - CATGTCAACGTCAAAGTCCAAG- 3’ R: 5’ - TGTGCACCCTCTGGTTAAAG - 3’
vCARD Caspase Recruitment domain (64R) – immediate early	F: 5’ - CGCGACCGTGATGGATAAA - 3’ R: 5’ - GCCAGATGCGGATCATCTT - 3’
Perkinsea (parasite)	F: 5’ - GAACGACCGTCCTACCTTGG - 3’ R: 5’ - AGGCCTGCTTGAAACACTCT - 3’

Non-polymorphic MHC class Ib (XNC) and invariant TCR alpha rearrangements

XNCs and iVT cells
Vα6-Jα1.43	F: 5’ - GGAATTGCTGGTTAATAATTTGGGA - 3’ R: 5’ - AAATGTGAGTTTGTGGAACCCCC - 3’
Vα22-Jα1.32	F: 5’ - ATGTGGTGGACGGACATACAG - 3’ R: 5’ - TGGAACCAGCTCCAGCATTAG - 3’
Vα23-Jα1.3	F: 5’ - CGCGATAAACTCAAAGGAAGAATA - 3’ R: 5’ - CTTTCCATAGCCTATTCCAGTA - 3’
Vα40-Jα1.22	F: 5’ - TCCAGCCTACCATGGAAGAAG - 3’ R: 5’ - CATATTTCCATAGCCACCAGTACT - 3’
Vα41-Jα1.40	F: 5’ - CCCAGCCTCCCAAGAAGGAGGTC - 3’ R: 5’ - GGTAAGTTTATTCCAGCCTCCTGT - 3’
Vα45-Jα1.14	F: 5’ - TCCGTTAAAGAGAAGGATTCCCAG - 3’ R: 5’ - CTCCCAGCCACTACCAGAATAAG - 3’
Vα39 SET 1	F: 5’ - ATATCTATGTGCTCTGCGGG - 3’ R: 5’ - GAGGCGATACATTGAAGGTT - 3’
Vα39 SET 2	F: 5’ - CAGAATATCACACCGGAGGA - 3’ R: 5’ - TGGCTGTCACTGTCACTATT - 3’
XNC1	F: 5’ - CATTTGAGGTGTGGCAAAGG - 3’ R: 5’ - GCAGCTCACATGCAGATTTC - 3’
XNC1	F: 5’ - GGGCTTTCCCAGTATGCCATT - 3’ R: 5’ - CAGCCTGTGCCTTTGCCACACC - 3’