Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI17133
Trapped Gene
Wdr76 (ENSMUSG00000027242)
Vector Insertion
Chr 2: 121336662 - 121343440
Public Clones (sanger) (sanger) (cmhd) CMHD-GT_486B3-3 (cmhd) CMHD-GT_364A11-3 (cmhd)
(cmhd) CMHD-GT_258E01-3 (cmhd)
Private Clones OST448809 (lexicon) OST412647 (lexicon) OST388126 (lexicon) OST323784 (lexicon)
OST309357 (lexicon) OST285064 (lexicon) OST252522 (lexicon) OST243932 (lexicon)
OST240985 (lexicon) OST240535 (lexicon) OST219881 (lexicon) OST136489 (lexicon)
OST123942 (lexicon) OST64017 (lexicon) OST59971 (lexicon) OST46391 (lexicon)
OST37723 (lexicon)
Severity of mutation (?) Insertion after 24% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000684574 (Chr2:121336275..121336661 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAAGCAGCGACGAAGACAGT Chr2:121336630..121336649 60.2 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000684574 (Chr2:121336275..121336661 +)
Downstram Exon
ENSMUSE00000167289 (Chr2:121343441..121343533 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAAGCAGCGACGAAGACAGT Chr2:121336630..121336649 60.2 50 TCAGCTAACTGAAGGGAAGCA Chr2:121343534..121343554 60.14 47.62

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000641680 Chr2:121332459..121332599 GATCGCTTAAAGGGCTAGGG Chr2:121332470..121332489 60.18 55
upstream ENSMUSE00000395929 Chr2:121332519..121332584 CTGGAAGGTCCTCCTGTCTG Chr2:121332542..121332561 59.83 60
upstream ENSMUSE00000684575 Chr2:121335584..121335671 GGTCCAAGGCTGAATCTGAG Chr2:121335616..121335635 59.8 55
upstream ENSMUSE00000167299 Chr2:121336275..121336661 AAAGCAGCGACGAAGACAGT Chr2:121336630..121336649 60.2 50
upstream ENSMUSE00000684574 Chr2:121336275..121336661 AAAGCAGCGACGAAGACAGT Chr2:121336630..121336649 60.2 50

*** Putative Vector Insertion (Chr 2: 121336662 - 121343440) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000167289 Chr2:121343441..121343533 TCAGCTAACTGAAGGGAAGCA Chr2:121343534..121343554 60.14 47.62
downstream ENSMUSE00000641678 Chr2:121343954..121344001 No primer for this exon
downstream ENSMUSE00000397606 Chr2:121345154..121345209 AGGTGACTCGCGCTTCTTTA Chr2:121345201..121345220 60.15 50
downstream ENSMUSE00000354245 Chr2:121351188..121351317 ATGGACCTTCGACATCCAAT Chr2:121351238..121351257 59.22 45
downstream ENSMUSE00000167293 Chr2:121352538..121352639 CATCTCTGCCCATGTCTTCA Chr2:121352636..121352655 59.79 50
downstream ENSMUSE00000167287 Chr2:121352738..121352781 AGACAGCCCCTTCTTGGTCT Chr2:121352773..121352792 60.25 55
downstream ENSMUSE00000167291 Chr2:121354547..121354700 GGAGATTGCTCCTTTGGTCA Chr2:121354622..121354641 60.19 50
downstream ENSMUSE00000355947 Chr2:121357022..121357177 AACACAGCGCTGGAAAAATC Chr2:121357173..121357192 60.26 45
downstream ENSMUSE00000641674 Chr2:121357473..121357496 GGCTTCTTGGTCAGACCCTA Chr2:121357496..121357515 59.28 55
downstream ENSMUSE00000597537 Chr2:121359844..121360058 CCCACTAGCAGACTGGAGGA Chr2:121359917..121359936 60.4 60
downstream ENSMUSE00000597536 Chr2:121361160..121361312 CAATGCTTTTCGAGTGCTCA Chr2:121361248..121361267 60.13 45
downstream ENSMUSE00000597535 Chr2:121362843..121362896 GGGAAGATATGGAGCTGCTG Chr2:121362874..121362893 59.8 55
downstream ENSMUSE00000597534 Chr2:121368037..121370593 ATAATGGCGCAAGGTCAAAC Chr2:121369269..121369288 59.97 45
downstream ENSMUSE00000684555 Chr2:121368037..121370596 ATAATGGCGCAAGGTCAAAC Chr2:121369269..121369288 59.97 45
downstream ENSMUSE00000684561 Chr2:121368037..121368745 CACATTCTTTCCCGACTCGT Chr2:121368175..121368194 60.11 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CACCTCTAATCGCCTTGCAG Chr2:121336707..121336727 60.92 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTTTCCTGTGTGGGGTTAGC Chr2:121336680..121336700 59.59 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027242