Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI15801
Trapped Gene
D230025D16Rik (ENSMUSG00000031889)
Vector Insertion
Chr 8: 107758466 - 107762322
Public Clones (sanger) (sanger) (ggtc) E058H07 (ggtc) D054A12 (ggtc) (ggtc)
E058H07 (ggtc) D054A12 (ggtc) E069E02 (ggtc) E046G10 (ggtc) IST13660B12 (tigm)
IST14641G2 (tigm) IST14513C6 (tigm) IST10730A4 (tigm) IST10107E6 (tigm)
IST12838F2 (tigm) IST14724A6 (tigm) IST14723E8 (tigm)
Private Clones OST472808 (lexicon) OST420930 (lexicon) OST330228 (lexicon) OST321903 (lexicon)
OST303242 (lexicon) OST235191 (lexicon) OST220573 (lexicon)
Severity of mutation (?) Insertion after 18% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000236942 (Chr8:107758382..107758465 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GACTCAGGACGGGATCACAC Chr8:107758414..107758433 60.54 60 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000236942 (Chr8:107758382..107758465 +)
Downstram Exon
ENSMUSE00000236921 (Chr8:107762323..107762366 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GACTCAGGACGGGATCACAC Chr8:107758414..107758433 60.54 60 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000405558 Chr8:107749099..107749386 CGAGCAATGGGAATTCACTC Chr8:107749364..107749383 60.6 50
upstream ENSMUSE00000236963 Chr8:107755028..107755110 AAGCACTGTCGCATCATCAG Chr8:107755063..107755082 60.02 50
upstream ENSMUSE00000236942 Chr8:107758382..107758465 GACTCAGGACGGGATCACAC Chr8:107758414..107758433 60.54 60

*** Putative Vector Insertion (Chr 8: 107758466 - 107762322) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000236921 Chr8:107762323..107762366 No primer for this exon
downstream ENSMUSE00000236900 Chr8:107763571..107763650 GAGCGATGGCCTGAGAGTTA Chr8:107763605..107763624 60.5 55
downstream ENSMUSE00000236883 Chr8:107763878..107763972 CCTCCGTCCACGAGTCTAAC Chr8:107763961..107763980 59.72 60
downstream ENSMUSE00000236858 Chr8:107764741..107764838 AAGCTAGACCATGGGCAAAA Chr8:107764768..107764787 59.71 45
downstream ENSMUSE00000236840 Chr8:107764927..107765039 GGTCCAGTTCCATCTCGAAG Chr8:107765010..107765029 59.65 55
downstream ENSMUSE00000214347 Chr8:107770321..107770454 CGCACGCTCAAATACTCTCA Chr8:107770376..107770395 60.16 50
downstream ENSMUSE00000214340 Chr8:107770738..107770821 GTTGCACTTGGACGGAACTT Chr8:107770791..107770810 60.16 50
downstream ENSMUSE00000214342 Chr8:107771996..107772078 TGCCCAGGGTAATTGGTATG Chr8:107772066..107772085 60.58 50
downstream ENSMUSE00000214344 Chr8:107772643..107772686 TGAACTCACAGCGGTGGTAA Chr8:107772665..107772684 60.3 50
downstream ENSMUSE00000214348 Chr8:107773319..107773362 CAGTCTGACCACCTGCATTTT Chr8:107773341..107773361 60.16 47.62
downstream ENSMUSE00000214346 Chr8:107773497..107773555 GCAAGACAACAGGCTTCTCC Chr8:107773554..107773573 60 55
downstream ENSMUSE00000214341 Chr8:107774619..107774691 GCCAAACGGATTGGTGTTAT Chr8:107774655..107774674 59.69 45
downstream ENSMUSE00000377433 Chr8:107775498..107776949 CTGTAGGCCCAGGAACCATA Chr8:107776741..107776760 59.95 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCTGTCTAATCGCCTTGCAG Chr8:107758511..107758531 59.17 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGCTGGGTGGATATGTTTCCT Chr8:107761421..107761442 59.84 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031889