Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI15643
Trapped Gene
Robo2 (ENSMUSG00000052516)
Vector Insertion
Chr 16: 73899156 - 73904802
Public Clones PST9026-NR (escells)
Private Clones not available
Severity of mutation (?) Insertion after 100% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000473218 (Chr16:73904628..73904801 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATAAGGGAGGACGGATGGAC Chr16:73904671..73904690 60.15 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000473218 (Chr16:73904628..73904801 -)
Downstram Exon
ENSMUSE00000475090 (Chr16:73899157..73899429 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATAAGGGAGGACGGATGGAC Chr16:73904671..73904690 60.15 55 CTGCTGCCTTAAACCCTGAC Chr16:73899358..73899377 59.88 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000467785 Chr16:74411097..74411157 TTGCTCTTTGGATTTCTCTGC Chr16:74411110..74411130 59.58 42.86
upstream ENSMUSE00000698695 Chr16:74411097..74412030 AGGAAACCAAGGGGAGAAAA Chr16:74411610..74411629 59.91 45
upstream ENSMUSE00000712927 Chr16:74411097..74411537 TGCTCTCTGGGATTGGTTTC Chr16:74411191..74411210 60.19 50
upstream ENSMUSE00000714056 Chr16:74411097..74411234 TGCTCTCTGGGATTGGTTTC Chr16:74411191..74411210 60.19 50
upstream ENSMUSE00000717868 Chr16:74411097..74412030 AGGAAACCAAGGGGAGAAAA Chr16:74411610..74411629 59.91 45
upstream ENSMUSE00000499472 Chr16:74352796..74353122 AGCCCACCACTCTGAACTGT Chr16:74353028..74353047 59.76 55
upstream ENSMUSE00000433345 Chr16:74047025..74047182 GAGATGACTTCCGGCAAAAC Chr16:74047157..74047176 59.68 50
upstream ENSMUSE00000698698 Chr16:74047025..74047186 GAGATGACTTCCGGCAAAAC Chr16:74047157..74047176 59.68 50
upstream ENSMUSE00000698713 Chr16:74047025..74047190 GAGATGACTTCCGGCAAAAC Chr16:74047157..74047176 59.68 50
upstream ENSMUSE00000433335 Chr16:74046067..74046187 GGAACCAATATGGTGGGAGA Chr16:74046102..74046121 59.6 50
upstream ENSMUSE00000698697 Chr16:74046067..74046187 GGAACCAATATGGTGGGAGA Chr16:74046102..74046121 59.6 50
upstream ENSMUSE00000506064 Chr16:74035230..74035368 ATTCCGTTGTCAGGTCCAAG Chr16:74035288..74035307 59.97 50
upstream ENSMUSE00000504033 Chr16:74016116..74016243 GAGAATCGGGTGGGAAAAGT Chr16:74016145..74016164 60.31 50
upstream ENSMUSE00000698696 Chr16:74009020..74009031 No primer for this exon
upstream ENSMUSE00000642683 Chr16:73996818..73996829 No primer for this exon
upstream ENSMUSE00000710135 Chr16:73986178..73986189 No primer for this exon
upstream ENSMUSE00000493265 Chr16:73985833..73985957 AAGGCCGAACAGTGACATTC Chr16:73985893..73985912 60.12 50
upstream ENSMUSE00000506606 Chr16:73982226..73982397 TTTTCCCGAATCAACCTCAG Chr16:73982371..73982390 60.04 45
upstream ENSMUSE00000512348 Chr16:73978653..73978858 TTCTGGGGAGAGATCCAAGA Chr16:73978698..73978717 59.73 50
upstream ENSMUSE00000513394 Chr16:73973950..73974031 GGAGTGCAGTGCTGGATGTA Chr16:73973954..73973973 59.86 55
upstream ENSMUSE00000511266 Chr16:73973398..73973560 GACCACCATCCAAACCTCAG Chr16:73973493..73973512 60.36 55
upstream ENSMUSE00000500556 Chr16:73971299..73971465 CGATCAACCCACAAGGTCTC Chr16:73971339..73971358 60.51 55
upstream ENSMUSE00000501579 Chr16:73968013..73968134 AGGAGTGGACCACAGACAGG Chr16:73968096..73968115 60.15 60
upstream ENSMUSE00000484042 Chr16:73961924..73962155 CGAAAACAGTCCGAACCACT Chr16:73961931..73961950 60.15 50
upstream ENSMUSE00000488535 Chr16:73958549..73958673 TCACAACAGCACAAGCATCA Chr16:73958608..73958627 60.03 45
upstream ENSMUSE00000487529 Chr16:73956733..73956904 ACGAAACGCGATTCCATATC Chr16:73956869..73956888 59.93 45
upstream ENSMUSE00000498042 Chr16:73948448..73948630 CACTGAGCAAATCACGGATG Chr16:73948568..73948587 60.26 50
upstream ENSMUSE00000495739 Chr16:73940501..73940543 No primer for this exon
upstream ENSMUSE00000494727 Chr16:73938976..73939103 CTTCTAAATGCTGGCGATCC Chr16:73939074..73939093 59.81 50
upstream ENSMUSE00000485740 Chr16:73933832..73934113 CTCTCGGATGGAGCCATTTA Chr16:73934047..73934066 60.17 50
upstream ENSMUSE00000698702 Chr16:73933291..73933416 ATTTCAGCACCACCAGCTCT Chr16:73933325..73933344 59.87 50
upstream ENSMUSE00000470989 Chr16:73928269..73928422 CACTATGCTGCGGAACAAGA Chr16:73928275..73928294 60.01 50
upstream ENSMUSE00000468283 Chr16:73920779..73921039 ACCATTACCGGTGCAAACAT Chr16:73920996..73921015 60.12 45
upstream ENSMUSE00000698686 Chr16:73920451..73921039 ACCATTACCGGTGCAAACAT Chr16:73920996..73921015 60.12 45
upstream ENSMUSE00000469188 Chr16:73916343..73916548 CAATACCCCACCTCCACAAC Chr16:73916517..73916536 60.09 55
upstream ENSMUSE00000473218 Chr16:73904628..73904801 ATAAGGGAGGACGGATGGAC Chr16:73904671..73904690 60.15 55
upstream ENSMUSE00000475090 Chr16:73899157..73899429 GTCAGGGTTTAAGGCAGCAG Chr16:73899380..73899399 59.88 55

*** Putative Vector Insertion (Chr 16: 73899156 - 73904802) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000698701 Chr16:73897261..73897452 GAAGGTCATTGGCATTCCTC Chr16:73897284..73897303 59.49 50
downstream ENSMUSE00000463747 Chr16:73897259..73897452 GAAGGTCATTGGCATTCCTC Chr16:73897284..73897303 59.49 50
downstream ENSMUSE00000698691 Chr16:73897252..73897452 GAAGGTCATTGGCATTCCTC Chr16:73897284..73897303 59.49 50
downstream ENSMUSE00000709629 Chr16:73894600..73896075 AGCCGAGTATGCTCGAAGAA Chr16:73894990..73895009 60.12 50
downstream ENSMUSE00000698700 Chr16:73892551..73896075 GATTTTGCGGGAATCACAGT Chr16:73893594..73893613 59.94 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AAGCTGTGTAATCGCCTTGC Chr16:73901739..73901759 60.42 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGCTGGAACTCTACCTGAACA Chr16:73901807..73901828 59.34 52.38 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

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