Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI5466
Trapped Gene
Grit (ENSMUSG00000041444)
Vector Insertion
Chr 9: 32015851 - 32046846
Public Clones CSJ270 (baygenomics) XG754 (baygenomics) (ggtc) (ggtc)
A051C04 (ggtc) (ggtc) (ggtc) A058A04 (ggtc) (ggtc) A058F01 (ggtc)
Private Clones OST423401 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000430641 (Chr9:32015785..32015850 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACCAACTCAGTGCCAAAACC Chr9:32015829..32015848 60.01 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000430641 (Chr9:32015785..32015850 +)
Downstram Exon
ENSMUSE00000390751 (Chr9:32046847..32046996 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACCAACTCAGTGCCAAAACC Chr9:32015829..32015848 60.01 50 AGTAATGAGCTTGCCGTGCT Chr9:32046878..32046897 60.04 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000430641 Chr9:32015785..32015850 ACCAACTCAGTGCCAAAACC Chr9:32015829..32015848 60.01 50

*** Putative Vector Insertion (Chr 9: 32015851 - 32046846) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000390751 Chr9:32046847..32046996 AGTAATGAGCTTGCCGTGCT Chr9:32046878..32046897 60.04 50
downstream ENSMUSE00000261059 Chr9:32052737..32052839 GCCATACCTCTCGATGAACG Chr9:32052786..32052805 60.62 55
downstream ENSMUSE00000261045 Chr9:32053503..32053647 TAGGTGAGCAGAGGGTTTGG Chr9:32053628..32053647 60.25 55
downstream ENSMUSE00000261021 Chr9:32054032..32054114 GTTGCTGGATGACATCATGG Chr9:32054098..32054117 59.92 50
downstream ENSMUSE00000375168 Chr9:32054693..32054800 CCACACAATGGCAAGGTTTT Chr9:32054786..32054805 60.79 45
downstream ENSMUSE00000260939 Chr9:32055071..32055216 CACTTCCATGAAAGCTGCTG Chr9:32055128..32055147 59.59 50
downstream ENSMUSE00000260921 Chr9:32055965..32056160 CTTGGGCCTTGACAGAGAAG Chr9:32055987..32056006 59.98 55
downstream ENSMUSE00000260902 Chr9:32058227..32058372 TTGCGTTTGGAGACAGAAGA Chr9:32058322..32058341 59.57 45
downstream ENSMUSE00000260879 Chr9:32061652..32061723 TTTAGCTGAGCGGAGGGTTC Chr9:32061689..32061708 61.76 55
downstream ENSMUSE00000260854 Chr9:32062759..32063643 GGGGCAGATTGTCATAGGAA Chr9:32062875..32062894 59.89 50
downstream ENSMUSE00000260823 Chr9:32064298..32065283 GGTGGGGTTTATCCACACTG Chr9:32064948..32064967 60.09 55
downstream ENSMUSE00000413669 Chr9:32066487..32068869 CATCAGGCCTAGCAAAGGAG Chr9:32066916..32066935 59.97 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ACTGTGCCTCTCTGGGAATG Chr9:32042842..32042862 60.26 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACTGTGCCTCTCTGGGAATG Chr9:32042842..32042862 60.26 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000041444