Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31197
Trapped Gene
Pcca (ENSMUSG00000041650)
Vector Insertion
Chr 14: 122962210 - 122981838
Public Clones IST14876D11 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000225114 (Chr14:122962141..122962209 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000225114 (Chr14:122962141..122962209 +)
Downstram Exon
ENSMUSE00000225109 (Chr14:122981839..122981952 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon GCTTTTACTGGTGGGAGCTG Chr14:122981901..122981920 59.88 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000361975 Chr14:122933561..122933681 No primer for this exon
upstream ENSMUSE00000225127 Chr14:122956937..122957014 TGCCTAGTGGTGTCCAGAAG Chr14:122956961..122956980 58.88 55
upstream ENSMUSE00000225122 Chr14:122962001..122962048 No primer for this exon
upstream ENSMUSE00000225114 Chr14:122962141..122962209 No primer for this exon

*** Putative Vector Insertion (Chr 14: 122962210 - 122981838) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000225109 Chr14:122981839..122981952 GCTTTTACTGGTGGGAGCTG Chr14:122981901..122981920 59.88 55
downstream ENSMUSE00000225101 Chr14:122984279..122984332 No primer for this exon
downstream ENSMUSE00000225095 Chr14:123015976..123016107 TGTCACCCATGGCTTGAATA Chr14:123016039..123016058 59.92 45
downstream ENSMUSE00000225087 Chr14:123037603..123037639 TTTCCCTTGCAATTCTGACA Chr14:123037639..123037658 59.25 40
downstream ENSMUSE00000225080 Chr14:123049474..123049552 No primer for this exon
downstream ENSMUSE00000225074 Chr14:123057900..123058002 CAGCTTCCTGGGATGAAAAT Chr14:123057934..123057953 59.13 45
downstream ENSMUSE00000225070 Chr14:123063476..123063570 CCTTCTCTGGATCGAGCACT Chr14:123063544..123063563 59.56 55
downstream ENSMUSE00000225065 Chr14:123067743..123067893 TTCCACAGTTCCAGCAGAAG Chr14:123067839..123067858 59.01 50
downstream ENSMUSE00000225060 Chr14:123084104..123084247 CAGCATAAACCCGACATTCA Chr14:123084248..123084267 59.54 45
downstream ENSMUSE00000225052 Chr14:123085325..123085399 CAGCCTCCCAATAGACGGTA Chr14:123085369..123085388 60.09 55
downstream ENSMUSE00000225042 Chr14:123089321..123089389 TTGGATGCCACTGTCAACTC Chr14:123089347..123089366 59.68 50
downstream ENSMUSE00000225033 Chr14:123091402..123091477 AATTGTCCAGTGCGTCTTCC Chr14:123091468..123091487 60.12 50
downstream ENSMUSE00000225026 Chr14:123100277..123100387 GAGCAATGGGATGTTGTGTG Chr14:123100305..123100324 59.97 50
downstream ENSMUSE00000225018 Chr14:123113245..123113347 TGGGATGCCACAAATACAGA Chr14:123113317..123113336 59.92 45
downstream ENSMUSE00000225011 Chr14:123137124..123137226 ACCGAGAGCTCCCACTTAGC Chr14:123137171..123137190 60.92 60
downstream ENSMUSE00000225003 Chr14:123189578..123189676 GTTTCGAGCCATCAACTTCC Chr14:123189602..123189621 59.68 50
downstream ENSMUSE00000224995 Chr14:123212416..123212469 GTGCCAAGAAACTGGATGCT Chr14:123212468..123212487 60.26 50
downstream ENSMUSE00000224991 Chr14:123275986..123276126 TGCAGCAAGCTTGGTTAAAA Chr14:123276021..123276040 59.62 40
downstream ENSMUSE00000224986 Chr14:123286269..123286346 CCATTTTCCCAGCTGTCATAC Chr14:123286343..123286363 59.44 47.62
downstream ENSMUSE00000479827 Chr14:123288709..123289166 GGAGACGCCAGTTAAAGCAG Chr14:123288832..123288851 60.01 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCTGTATTCCCCAAATGGAT Chr14:122962220..122962240 58.72 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CGTCATACGTGACTGGGAAAA Chr14:122962254..122962275 60.93 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000041650