Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32935
Trapped Gene
Lrriq4 (ENSMUSG00000027703)
Vector Insertion
Chr 3: 30544271 - 30545174
Public Clones IST12045D10 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000639711 (Chr3:30544140..30544270 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGGAGCTTCTCCATCGACT Chr3:30544212..30544231 59.56 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000639711 (Chr3:30544140..30544270 +)
Downstram Exon
ENSMUSE00000172454 (Chr3:30545175..30545235 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGGAGCTTCTCCATCGACT Chr3:30544212..30544231 59.56 55 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000676165 Chr3:30543429..30543774 CAAGAGCAACAACCAGGACA Chr3:30543716..30543735 59.87 50
upstream ENSMUSE00000639711 Chr3:30544140..30544270 CTGGAGCTTCTCCATCGACT Chr3:30544212..30544231 59.56 55

*** Putative Vector Insertion (Chr 3: 30544271 - 30545174) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000172454 Chr3:30545175..30545235 No primer for this exon
downstream ENSMUSE00000676162 Chr3:30547148..30547182 No primer for this exon
downstream ENSMUSE00000274098 Chr3:30548827..30549874 ATCTGCACGGGAAAGTGTTC Chr3:30549822..30549841 60.12 50
downstream ENSMUSE00000172455 Chr3:30554559..30554732 ATTTCCTGTGAGCCCGAGTA Chr3:30554618..30554637 59.69 50
downstream ENSMUSE00000172449 Chr3:30557976..30558114 GCTGTTCCAGTCGGTTGTTC Chr3:30558021..30558040 60.7 55
downstream ENSMUSE00000172451 Chr3:30558823..30559019 CATGGGGTTGTTCATCAGTG Chr3:30558929..30558948 59.81 50
downstream ENSMUSE00000676164 Chr3:30571063..30571353 GTCCTTTGGAGGGGATTTTC Chr3:30571164..30571183 59.74 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCCATCGACTCTGGAGCTTT Chr3:30544223..30544243 59.95 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCCATCGACTCTGGAGCTTT Chr3:30544223..30544243 59.95 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027703