Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6562
Trapped Gene
Fbxo21 (ENSMUSG00000032898)
Vector Insertion
Chr 5: 118452182 - 118458018
Public Clones YHB304 (baygenomics) YHA151 (baygenomics) YHB409 (baygenomics) YHB116 (baygenomics)
YHB417 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 90% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000291132 (Chr5:118452024..118452181 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGAGTGGATCCGAAACATGA Chr5:118452078..118452097 60.05 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000291132 (Chr5:118452024..118452181 +)
Downstram Exon
ENSMUSE00000385255 (Chr5:118458019..118460199 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGAGTGGATCCGAAACATGA Chr5:118452078..118452097 60.05 45 CCGAGGCTGCTTAGTACTGG Chr5:118459751..118459770 60.03 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000380120 Chr5:118426801..118427048 TGCTGGAGTACATCCTGTGC Chr5:118426919..118426938 59.86 55
upstream ENSMUSE00000291191 Chr5:118427867..118428002 CCAAGCGGTTCTTTTCAGAG Chr5:118427980..118427999 59.99 50
upstream ENSMUSE00000291184 Chr5:118429750..118429844 GAGGGCCCAGAGATCTTTTT Chr5:118429786..118429805 59.65 50
upstream ENSMUSE00000291179 Chr5:118436108..118436229 CAATAACCTCAAGGCGTTCC Chr5:118436174..118436193 59.57 50
upstream ENSMUSE00000291172 Chr5:118438787..118438933 AACCCACTCTCCGACATCAG Chr5:118438813..118438832 60.11 55
upstream ENSMUSE00000291166 Chr5:118439133..118439269 CAAGTGCTGGACGCTATCAA Chr5:118439171..118439190 60.01 50
upstream ENSMUSE00000291159 Chr5:118440812..118440948 AACTTCCCAAGCCACTTCCT Chr5:118440902..118440921 60.11 50
upstream ENSMUSE00000291151 Chr5:118444491..118444670 AGAAGGTGCTACAGCGGATG Chr5:118444622..118444641 60.42 55
upstream ENSMUSE00000291145 Chr5:118445379..118445511 TCTACTTCCACCTGGGCATC Chr5:118445480..118445499 60.07 55
upstream ENSMUSE00000291138 Chr5:118450326..118450537 AGGTGCTTGACATCCTCCAG Chr5:118450345..118450364 60.26 55
upstream ENSMUSE00000291132 Chr5:118452024..118452181 TGAGTGGATCCGAAACATGA Chr5:118452078..118452097 60.05 45

*** Putative Vector Insertion (Chr 5: 118452182 - 118458018) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000385255 Chr5:118458019..118460199 CCGAGGCTGCTTAGTACTGG Chr5:118459751..118459770 60.03 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAGATATGCAGCCCAAGGTAA Chr5:118452166..118452187 60.1 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAGATATGCAGCCCAAGGTAA Chr5:118452166..118452187 60.1 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032898