Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI36665
Trapped Gene
Ucp3 (ENSMUSG00000032942)
Vector Insertion
Chr 7: 107631265 - 107633502
Public Clones IST12449D9 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 88% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000225886 (Chr7:107631084..107631264 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGCCCTCTGCACTGTATGCT Chr7:107631203..107631222 60.04 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000225886 (Chr7:107631084..107631264 +)
Downstram Exon
ENSMUSE00000671973 (Chr7:107633503..107633815 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGCCCTCTGCACTGTATGCT Chr7:107631203..107631222 60.04 55 GGACGAAACACGGAGGACTA Chr7:107633761..107633780 60.11 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000225926 Chr7:107621500..107621603 TACCCAACCTTGGCTAGACG Chr7:107621579..107621598 60.12 55
upstream ENSMUSE00000671975 Chr7:107621502..107621603 TACCCAACCTTGGCTAGACG Chr7:107621579..107621598 60.12 55
upstream ENSMUSE00000225915 Chr7:107627883..107628101 AAGCTCTCTCCTCGGACCTC Chr7:107627918..107627937 60.1 60
upstream ENSMUSE00000671974 Chr7:107627943..107628101 CAACGGTTGTGAAGTTCCTG Chr7:107628010..107628029 59.18 50
upstream ENSMUSE00000202247 Chr7:107628595..107628796 CGAATTGGCCTCTACGACTC Chr7:107628742..107628761 59.84 55
upstream ENSMUSE00000202239 Chr7:107628990..107629190 GTCCGATTTCAAGCCATGAT Chr7:107629076..107629095 59.9 45
upstream ENSMUSE00000225897 Chr7:107630380..107630481 TGCTGAGATGGTGACCTACG Chr7:107630420..107630439 59.85 55
upstream ENSMUSE00000225886 Chr7:107631084..107631264 AGCCCTCTGCACTGTATGCT Chr7:107631203..107631222 60.04 55

*** Putative Vector Insertion (Chr 7: 107631265 - 107633502) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000348151 Chr7:107633503..107634940 GTATAGGGCGCTCAAATGGA Chr7:107634867..107634886 60.06 50
downstream ENSMUSE00000671973 Chr7:107633503..107633815 GGACGAAACACGGAGGACTA Chr7:107633761..107633780 60.11 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGTAAGCTGTCACCCTCTGC Chr7:107631265..107631285 59.87 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGTAAGCTGTCACCCTCTGC Chr7:107631265..107631285 59.87 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032942