Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI28195
Trapped Gene
Ucp2 (ENSMUSG00000033685)
Vector Insertion
Chr 7: 107641975 - 107642722
Public Clones IST14022F11 (tigm)
Private Clones OST37562 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000390980 (Chr7:107641880..107641974 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACAGCCTTCTGCACTCCTGT Chr7:107641939..107641958 60.06 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000390980 (Chr7:107641880..107641974 +)
Downstram Exon
ENSMUSE00000202263 (Chr7:107642723..107642884 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACAGCCTTCTGCACTCCTGT Chr7:107641939..107641958 60.06 55 CCTCCAAGGTCAAGCTTCTCT Chr7:107642855..107642875 60 52.38

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

*** Putative Vector Insertion (Chr 7: 107641975 - 107642722) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000202263 Chr7:107642723..107642884 CCTCCAAGGTCAAGCTTCTCT Chr7:107642855..107642875 60 52.38
downstream ENSMUSE00000202236 Chr7:107645235..107645457 ACATCTGTGGCCTTGAAACC Chr7:107645360..107645379 59.97 50
downstream ENSMUSE00000202257 Chr7:107645604..107645814 ATTGTAGAGGCTGCGTGGAC Chr7:107645720..107645739 60.28 55
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 ACACAGCCTTCTGCACTCCT Chr7:107641938..107641958 60.06 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACACAGCCTTCTGCACTCCT Chr7:107641938..107641958 60.06 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000033685