Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI705
Trapped Gene
Pggt1b (ENSMUSG00000024477)
Vector Insertion
Chr 18: 46408710 - 46413932
Public Clones DB0211 (sanger) IST14889D12 (tigm) IST14885D12 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 58% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000142720 (Chr18:46413933..46413978 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000142720 (Chr18:46413933..46413978 -)
Downstram Exon
ENSMUSE00000142716 (Chr18:46408525..46408709 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon TTACCCATCAGGCACAGTGA Chr18:46408642..46408661 60.11 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000367844 Chr18:46440334..46440504 AAGGAGAACGGCTGGATTTC Chr18:46440414..46440433 60.58 50
upstream ENSMUSE00000142722 Chr18:46434244..46434362 TCCTTGGATGTGGTGAACAA Chr18:46434297..46434316 59.94 45
upstream ENSMUSE00000142709 Chr18:46422544..46422611 GTGGTTTCCGAGGTTCTTCA Chr18:46422574..46422593 60.09 50
upstream ENSMUSE00000142723 Chr18:46419258..46419409 AGATGACTTAGGCCGTGTGG Chr18:46419310..46419329 60.13 55
upstream ENSMUSE00000142718 Chr18:46417739..46417871 TTTGTGTACTGCGCATCCTG Chr18:46417812..46417831 60.87 50
upstream ENSMUSE00000142720 Chr18:46413933..46413978 No primer for this exon

*** Putative Vector Insertion (Chr 18: 46408710 - 46413932) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000142716 Chr18:46408525..46408709 TTACCCATCAGGCACAGTGA Chr18:46408642..46408661 60.11 50
downstream ENSMUSE00000142710 Chr18:46406216..46406324 CCAACAAGGCGATCTTGAGT Chr18:46406226..46406245 60.26 50
downstream ENSMUSE00000379175 Chr18:46399603..46401355 TTCCAATTGTGTCACGCTGT Chr18:46399943..46399962 60.16 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCCTGTTAGCACTGTTTGGA Chr18:46410898..46410919 59.93 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTACCGTGACTGGGAAAACC Chr18:46410865..46410885 59.83 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CTCATGGTATAATCGCCTTGC Chr18:46410916..46410937 59.58 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 ACAAGGTCTTCTCGTGACTGG Chr18:46413919..46413940 59.36 52.38 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024477