Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI21943
Trapped Gene
Gm920 (ENSMUSG00000075413)
Vector Insertion
Chr 15: 99785352 - 99785873
Public Clones CMHD-GT_490E8-3 (cmhd)
Private Clones OST313453 (lexicon) OST210821 (lexicon)
Severity of mutation (?) Insertion after 16% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000646075 (Chr15:99785874..99786090 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TAATGAGCAACGTGGAGCTG Chr15:99786046..99786065 60.01 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000646075 (Chr15:99785874..99786090 -)
Downstram Exon
ENSMUSE00000646074 (Chr15:99785181..99785351 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TAATGAGCAACGTGGAGCTG Chr15:99786046..99786065 60.01 50 ATCCTTTTCAGCGTGTCTGG Chr15:99785236..99785255 60.26 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000646076 Chr15:99797181..99797492 TGTAACCTCGGGAGTCCTTG Chr15:99797464..99797483 60.1 55
upstream ENSMUSE00000646075 Chr15:99785874..99786090 TAATGAGCAACGTGGAGCTG Chr15:99786046..99786065 60.01 50

*** Putative Vector Insertion (Chr 15: 99785352 - 99785873) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000646074 Chr15:99785181..99785351 ATCCTTTTCAGCGTGTCTGG Chr15:99785236..99785255 60.26 50
downstream ENSMUSE00000646072 Chr15:99776152..99778206 CATTACACCAGGCTCCACCT Chr15:99776365..99776384 59.99 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTCCTGGCTAGAAGAATGGA Chr15:99785872..99785893 59.82 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGGAAGGAGCAAAGTTAAGGA Chr15:99785828..99785849 59.71 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000075413