Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14425
Trapped Gene
Psmd14 (ENSMUSG00000026914)
Vector Insertion
Chr 2: 61598765 - 61599031
Public Clones FHCRC-GT-S13-4D1 (fhcrc)
Private Clones not available
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000350608 (Chr2:61598693..61598764 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TCTTCCTTGGCCTTGCTAAA Chr2:61598744..61598763 59.95 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000350608 (Chr2:61598693..61598764 +)
Downstram Exon
ENSMUSE00000164026 (Chr2:61599032..61599151 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TCTTCCTTGGCCTTGCTAAA Chr2:61598744..61598763 59.95 45 TCCAGCACGACCATGTTTTA Chr2:61599058..61599077 60.11 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

*** Putative Vector Insertion (Chr 2: 61598765 - 61599031) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000164026 Chr2:61599032..61599151 TCCAGCACGACCATGTTTTA Chr2:61599058..61599077 60.11 45
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 AAAGGTAAGGAAGGGGTGGA Chr2:61598762..61598782 59.8 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCTTCCTTTGCATTTGCTTC Chr2:61598797..61598817 59.97 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026914