Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI8064
Trapped Gene
Psmd14 (ENSMUSG00000026914)
Vector Insertion
Chr 2: 61599152 - 61601893
Public Clones RRJ236 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 26% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000164026 (Chr2:61599032..61599151 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TAAAACATGGTCGTGCTGGA Chr2:61599036..61599055 60.11 45 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000164026 (Chr2:61599032..61599151 +)
Downstram Exon
ENSMUSE00000164029 (Chr2:61601894..61601964 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TAAAACATGGTCGTGCTGGA Chr2:61599036..61599055 60.11 45 CCAACATTTTGGCTTGGAAC Chr2:61601945..61601964 60.34 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000164032 Chr2:61549816..61549930 GCTGATTCATCGCCTGTTCT Chr2:61549849..61549868 60.37 50
upstream ENSMUSE00000164024 Chr2:61558501..61558628 No primer for this exon
upstream ENSMUSE00000386329 Chr2:61560767..61560818 GAGGAGGTATGCCTGGACTG Chr2:61560793..61560812 59.68 60
upstream ENSMUSE00000350608 Chr2:61598693..61598764 TCTTCCTTGGCCTTGCTAAA Chr2:61598744..61598763 59.95 45
upstream ENSMUSE00000164026 Chr2:61599032..61599151 TAAAACATGGTCGTGCTGGA Chr2:61599036..61599055 60.11 45

*** Putative Vector Insertion (Chr 2: 61599152 - 61601893) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000164029 Chr2:61601894..61601964 CCAACATTTTGGCTTGGAAC Chr2:61601945..61601964 60.34 45
downstream ENSMUSE00000164030 Chr2:61602906..61603056 GAATGGGATCCACAACCACT Chr2:61603042..61603061 59.64 50
downstream ENSMUSE00000164022 Chr2:61614730..61614837 GCTTGTTTAAGTGGCCCAGA Chr2:61614829..61614848 60.25 50
downstream ENSMUSE00000164034 Chr2:61621890..61621964 No primer for this exon
downstream ENSMUSE00000164018 Chr2:61623491..61623616 CCTGAAGTGTCAATCCTTCCA Chr2:61623544..61623564 60.1 47.62
downstream ENSMUSE00000164020 Chr2:61635452..61635514 CCAGCTGTTCAGGTGTCATC Chr2:61635494..61635513 59.26 55
downstream ENSMUSE00000341036 Chr2:61638039..61638432 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGCTATGCCACAGTCAGGAA Chr2:61599131..61599151 60.41 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGCTATGCCACAGTCAGGAA Chr2:61599131..61599151 60.41 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026914