Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI18639
Trapped Gene
Gla (ENSMUSG00000031266)
Vector Insertion
Chr X: 131130989 - 131135276
Public Clones IST12452G12 (tigm)
Private Clones OST418880 (lexicon) OST80371 (lexicon)
Severity of mutation (?) Insertion after 16% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000352917 (ChrX:131135277..131135494 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGCATTCTTGGGGTCAGAG ChrX:131135380..131135399 59.8 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000352917 (ChrX:131135277..131135494 -)
Downstram Exon
ENSMUSE00000207762 (ChrX:131130814..131130988 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGCATTCTTGGGGTCAGAG ChrX:131135380..131135399 59.8 55 ATTTGCGAGGTGTTTGATCC ChrX:131130795..131130814 59.94 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000352917 ChrX:131135277..131135494 GAGCATTCTTGGGGTCAGAG ChrX:131135380..131135399 59.8 55

*** Putative Vector Insertion (Chr X: 131130989 - 131135276) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000207762 ChrX:131130814..131130988 ATTTGCGAGGTGTTTGATCC ChrX:131130795..131130814 59.94 45
downstream ENSMUSE00000277094 ChrX:131129719..131129896 TAGCAGATCTACGCCCCAGT ChrX:131129743..131129762 59.86 55
downstream ENSMUSE00000207760 ChrX:131128173..131128264 ATACAATGCTTCGGCCTGTC ChrX:131128192..131128211 60.1 50
downstream ENSMUSE00000207767 ChrX:131126604..131126765 TCCAGCGACTTCAACAATCTC ChrX:131126609..131126629 60.39 47.62
downstream ENSMUSE00000207764 ChrX:131125485..131125682 GCAGAGCTTTGGCTTGAGAG ChrX:131125531..131125550 60.42 55
downstream ENSMUSE00000277059 ChrX:131122895..131124666 TCTCGGGGAGAAGCTGAGTA ChrX:131124473..131124492 60.09 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTTGTGTGGTAATCGCCTTG ChrX:131135214..131135234 59.58 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCTTGACTGCCAAGAAGAGC ChrX:131135291..131135311 60.13 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 ATACGCTAATCGCCTTGCAG ChrX:131135430..131135450 60.38 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CTGCGCTTTGTCATCTTTTT ChrX:131135524..131135544 58.17 40 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031266