Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI26556
Trapped Gene
Mto1 (ENSMUSG00000032342)
Vector Insertion
Chr 9: 78296318 - 78297225
Public Clones D177B08 (ggtc)
Private Clones OST111032 (lexicon)
Severity of mutation (?) Insertion after 11% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000218877 (Chr9:78296027..78296317 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGGCTTCTACTCCGGTGAC Chr9:78296051..78296070 59.87 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000218877 (Chr9:78296027..78296317 +)
Downstram Exon
ENSMUSE00000218893 (Chr9:78297226..78297425 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGGCTTCTACTCCGGTGAC Chr9:78296051..78296070 59.87 60 AGTTTCCGGTCAATCTGTGC Chr9:78297409..78297428 60.12 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000218877 Chr9:78296027..78296317 CTGGCTTCTACTCCGGTGAC Chr9:78296051..78296070 59.87 60

*** Putative Vector Insertion (Chr 9: 78296318 - 78297225) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000218893 Chr9:78297226..78297425 AGTTTCCGGTCAATCTGTGC Chr9:78297409..78297428 60.12 50
downstream ENSMUSE00000353823 Chr9:78297525..78297642 GGTACCCAGGCTCTGGTTCT Chr9:78297612..78297631 60.51 60
downstream ENSMUSE00000269893 Chr9:78300571..78300860 GTCTGAGCCAACCCTATGGA Chr9:78300709..78300728 60.07 55
downstream ENSMUSE00000218895 Chr9:78305017..78305129 AAGGACAATCGCATCCACTC Chr9:78305082..78305101 60.08 50
downstream ENSMUSE00000269839 Chr9:78305223..78305413 CTGATGCAGACGATTTGGAA Chr9:78305280..78305299 59.8 45
downstream ENSMUSE00000269816 Chr9:78305724..78305854 CTGATCTGACGGGGGTCTAA Chr9:78305769..78305788 60.06 55
downstream ENSMUSE00000218875 Chr9:78308640..78308844 GTGGCTTTCGACTGACTCGT Chr9:78308691..78308710 60.45 55
downstream ENSMUSE00000218882 Chr9:78309327..78309498 TCAAATCGCTGTGAGGACAC Chr9:78309369..78309388 59.84 50
downstream ENSMUSE00000269774 Chr9:78312691..78312809 GAGCGACTCCATGTCCACTT Chr9:78312739..78312758 60.27 55
downstream ENSMUSE00000269759 Chr9:78318438..78318598 CTGTGGGCGACTCAAGTGTA Chr9:78318598..78318617 59.9 55
downstream ENSMUSE00000218867 Chr9:78321597..78321957 GAGACTCCCACAGGGATTGA Chr9:78321895..78321914 60.05 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr9:78296369..78296389 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector

Come back to gene ENSMUSG00000032342