Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI19398
Trapped Gene
Lap3 (ENSMUSG00000039682)
Vector Insertion
Chr 5: 45886521 - 45888158
Public Clones (sanger)
Private Clones OST392036 (lexicon)
Severity of mutation (?) Insertion after 14% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000287289 (Chr5:45886405..45886520 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAAGTGCGGGAGAGAGTTTT Chr5:45886460..45886479 59.47 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000287289 (Chr5:45886405..45886520 +)
Downstram Exon
ENSMUSE00000287283 (Chr5:45888159..45888213 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAAGTGCGGGAGAGAGTTTT Chr5:45886460..45886479 59.47 50 GCTTTGAGAGGAGGTCCAGA Chr5:45888182..45888201 59.53 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000287307 Chr5:45884699..45884800 TGGGAGTGAGAGGTCTCTGG Chr5:45884748..45884767 60.39 60
upstream ENSMUSE00000287289 Chr5:45886405..45886520 CAAGTGCGGGAGAGAGTTTT Chr5:45886460..45886479 59.47 50

*** Putative Vector Insertion (Chr 5: 45886521 - 45888158) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000287283 Chr5:45888159..45888213 GCTTTGAGAGGAGGTCCAGA Chr5:45888182..45888201 59.53 55
downstream ENSMUSE00000287276 Chr5:45888568..45888673 GCCAGTTTTCCTGGTCATCA Chr5:45888640..45888659 61.05 50
downstream ENSMUSE00000287270 Chr5:45889673..45889832 GAGACCAAGCACAGCTCCTT Chr5:45889773..45889792 59.6 55
downstream ENSMUSE00000287264 Chr5:45891642..45891806 ATTTCAGCGAATCTGGTTGG Chr5:45891758..45891777 60.07 45
downstream ENSMUSE00000287253 Chr5:45894619..45894777 GCTGCCCATGTAGTGGATTT Chr5:45894724..45894743 59.96 50
downstream ENSMUSE00000287241 Chr5:45895962..45896086 CATGTTTGCAGAAGCCTTGA Chr5:45895998..45896017 59.99 45
downstream ENSMUSE00000287235 Chr5:45897413..45897501 TTGCCGCTAGGCATATTTTC Chr5:45897452..45897471 60.19 45
downstream ENSMUSE00000287231 Chr5:45898346..45898448 CTCGGCGTCAGTGTTATCAA Chr5:45898369..45898388 59.86 50
downstream ENSMUSE00000287221 Chr5:45900677..45900756 AGCCAGGACGAATTGGTAAA Chr5:45900740..45900759 59.57 45
downstream ENSMUSE00000287216 Chr5:45902347..45902456 GACTCGGTCCCCAGTTTCTAC Chr5:45902375..45902395 59.99 57.14
downstream ENSMUSE00000467351 Chr5:45903120..45903394 TGTGCCCACTTCGTATGTGT Chr5:45903188..45903207 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGGGTTCCTTTTTGTTTGTG Chr5:45886540..45886560 59.31 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGGGTTCCTTTTTGTTTGTG Chr5:45886540..45886560 59.31 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000039682