Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33530
Trapped Gene
Ucp2 (ENSMUSG00000033685)
Vector Insertion
Chr 7: 107645815 - 107646586
Public Clones IST10170A11 (tigm) IST13132C8 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 36% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000202257 (Chr7:107645604..107645814 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTCCACGCAGCCTCTACAAT Chr7:107645698..107645717 60.28 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000202257 (Chr7:107645604..107645814 +)
Downstram Exon
ENSMUSE00000202264 (Chr7:107646587..107646781 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTCCACGCAGCCTCTACAAT Chr7:107645698..107645717 60.28 55 TCGTGCAATGGTCTTGTAGG Chr7:107646756..107646775 59.72 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000390980 Chr7:107641880..107641974 ACAGCCTTCTGCACTCCTGT Chr7:107641939..107641958 60.06 55
upstream ENSMUSE00000202263 Chr7:107642723..107642884 CTCCCAGCCATTTTCTATGG Chr7:107642754..107642773 59.52 50
upstream ENSMUSE00000202236 Chr7:107645235..107645457 ATTGAAGGTCCCCGTTTCTC Chr7:107645269..107645288 60.31 50
upstream ENSMUSE00000202257 Chr7:107645604..107645814 GTCCACGCAGCCTCTACAAT Chr7:107645698..107645717 60.28 55

*** Putative Vector Insertion (Chr 7: 107645815 - 107646586) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000202264 Chr7:107646587..107646781 TCGTGCAATGGTCTTGTAGG Chr7:107646756..107646775 59.72 50
downstream ENSMUSE00000202237 Chr7:107646857..107646958 TGTCATGAGGTTGGCTTTCA Chr7:107646960..107646979 60.24 45
downstream ENSMUSE00000202235 Chr7:107647245..107647425 GAGCATGGTAAGGGCACAGT Chr7:107647396..107647415 60.14 55
downstream ENSMUSE00000449447 Chr7:107647745..107648134 CATGGAGAGGCTCAGAAAGG Chr7:107647873..107647892 59.94 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTATGCGGCAAGGACAGACT Chr7:107645820..107645840 60.28 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTATGCGGCAAGGACAGACT Chr7:107645820..107645840 60.28 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000033685