Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI29440
Trapped Gene
1700112C13Rik (ENSMUSG00000049719)
Vector Insertion
Chr 9: 110752763 - 110753818
Public Clones (cmhd) CMHD-GT_410D11-3 (cmhd) IST13114F9 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 45% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000391531 (Chr9:110752500..110752762 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATCAATCGCATGTCTGACGA Chr9:110752606..110752625 60.23 45 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000391531 (Chr9:110752500..110752762 +)
Downstram Exon
ENSMUSE00000220137 (Chr9:110753819..110753982 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATCAATCGCATGTCTGACGA Chr9:110752606..110752625 60.23 45 GGAGGAACTGGTACCGATGA Chr9:110753908..110753927 59.93 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000583134 Chr9:110747010..110747138 CAGTGGATCCTCACGGTCTT Chr9:110747067..110747086 60.11 55
upstream ENSMUSE00000715741 Chr9:110752042..110752308 ACCTGCAAGGTGGTAAATGG Chr9:110752166..110752185 59.85 50
upstream ENSMUSE00000348778 Chr9:110752131..110752308 GCAAGGTGGTAAATGGGAAG Chr9:110752170..110752189 59.43 50
upstream ENSMUSE00000391531 Chr9:110752500..110752762 ATCAATCGCATGTCTGACGA Chr9:110752606..110752625 60.23 45

*** Putative Vector Insertion (Chr 9: 110752763 - 110753818) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000220137 Chr9:110753819..110753982 GGAGGAACTGGTACCGATGA Chr9:110753908..110753927 59.93 55
downstream ENSMUSE00000406701 Chr9:110758504..110759026 CATCTGGACCCAGGTCTTGT Chr9:110758560..110758579 59.96 55
downstream ENSMUSE00000715516 Chr9:110758504..110759020 CATCTGGACCCAGGTCTTGT Chr9:110758560..110758579 59.96 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CGAAGGTGAGTGCAGTAGAGC Chr9:110752759..110752780 60.21 57.14 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000049719