Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32145
Trapped Gene
Man1b1 (ENSMUSG00000036646)
Vector Insertion
Chr 2: 25197237 - 25198743
Public Clones IST12525A6 (tigm) IST11910F3 (tigm) IST12150G3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 31% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000242449 (Chr2:25197127..25197236 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAGAGCTGAAGCCTCCATCA Chr2:25197178..25197197 60.1 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000242449 (Chr2:25197127..25197236 +)
Downstram Exon
ENSMUSE00000715697 (Chr2:25198744..25198929 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAGAGCTGAAGCCTCCATCA Chr2:25197178..25197197 60.1 50 TTGCTTCAGACCCAAAATCC Chr2:25198931..25198950 60.05 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000698883 Chr2:25187935..25188102 CAAGCCAGACGTAACTGCAA Chr2:25187991..25188010 60.05 50
upstream ENSMUSE00000445160 Chr2:25188276..25188496 GGTGTGGATGCCTTCGTTAT Chr2:25188282..25188301 59.82 50
upstream ENSMUSE00000445144 Chr2:25188955..25189063 AGCAGCTTTCGAGGCTACAG Chr2:25188962..25188981 59.92 55
upstream ENSMUSE00000445140 Chr2:25189897..25190012 GTCTTACCAGCGCCTCAGAA Chr2:25189950..25189969 60.54 55
upstream ENSMUSE00000242453 Chr2:25193633..25193787 CAGTCTCCAAGGATGGGATG Chr2:25193691..25193710 60.47 55
upstream ENSMUSE00000242449 Chr2:25197127..25197236 AAGAGCTGAAGCCTCCATCA Chr2:25197178..25197197 60.1 50

*** Putative Vector Insertion (Chr 2: 25197237 - 25198743) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000714026 Chr2:25198744..25198929 TTGCTTCAGACCCAAAATCC Chr2:25198931..25198950 60.05 45
downstream ENSMUSE00000715697 Chr2:25198744..25198929 TTGCTTCAGACCCAAAATCC Chr2:25198931..25198950 60.05 45
downstream ENSMUSE00000242442 Chr2:25200488..25200636 CCTAAGATGCGAATGGTGCT Chr2:25200584..25200603 60.24 50
downstream ENSMUSE00000698872 Chr2:25200488..25200636 CCTAAGATGCGAATGGTGCT Chr2:25200584..25200603 60.24 50
downstream ENSMUSE00000242438 Chr2:25200926..25201114 CTCCAACTGAATGCTCGTCA Chr2:25201075..25201094 59.98 50
downstream ENSMUSE00000698871 Chr2:25200926..25201114 CTCCAACTGAATGCTCGTCA Chr2:25201075..25201094 59.98 50
downstream ENSMUSE00000242436 Chr2:25203556..25203746 CTGGATCCACTGTTTCAGCA Chr2:25203729..25203748 59.83 50
downstream ENSMUSE00000698870 Chr2:25203556..25203746 CTGGATCCACTGTTTCAGCA Chr2:25203729..25203748 59.83 50
downstream ENSMUSE00000242433 Chr2:25204091..25204211 AGCTTTCTGGGCTGAGACTG Chr2:25204168..25204187 59.74 55
downstream ENSMUSE00000698869 Chr2:25204091..25204211 AGCTTTCTGGGCTGAGACTG Chr2:25204168..25204187 59.74 55
downstream ENSMUSE00000242427 Chr2:25204690..25204887 CCGAGCTAGATCCATGTGGT Chr2:25204776..25204795 60.1 55
downstream ENSMUSE00000698868 Chr2:25204690..25204887 CCGAGCTAGATCCATGTGGT Chr2:25204776..25204795 60.1 55
downstream ENSMUSE00000242423 Chr2:25204971..25205102 TAGGCTTTCCACCGTCTCTG Chr2:25205021..25205040 60.39 55
downstream ENSMUSE00000698867 Chr2:25204971..25205102 TAGGCTTTCCACCGTCTCTG Chr2:25205021..25205040 60.39 55
downstream ENSMUSE00000380274 Chr2:25205882..25207732 CCCTTGACACCAGCCATAGT Chr2:25207515..25207534 59.99 55
downstream ENSMUSE00000698865 Chr2:25205882..25207732 CCCTTGACACCAGCCATAGT Chr2:25207515..25207534 59.99 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AAGAGCCTGTAATCGCCTTG Chr2:25197279..25197299 59.48 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCCCTTACCTCACCAGAACC Chr2:25197252..25197272 60.74 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000036646