Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI5866
Trapped Gene
Zfyve19 (ENSMUSG00000068580)
Vector Insertion
Chr 2: 119037036 - 119037133
Public Clones CSH950 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 30% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000562406 (Chr2:119036917..119037035 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCTTCGTCAGGAAAACAAGC Chr2:119037013..119037032 60 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000562406 (Chr2:119036917..119037035 +)
Downstram Exon
ENSMUSE00000562405 (Chr2:119037134..119037279 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCTTCGTCAGGAAAACAAGC Chr2:119037013..119037032 60 50 CTAGCCGTGCCTCTATCTCG Chr2:119037175..119037194 60.13 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000661811 Chr2:119034761..119034919 GCCGAGAACTGCTATGGAGA Chr2:119034853..119034872 60.5 55
upstream ENSMUSE00000562408 Chr2:119036218..119036339 CTTAGCTTCAGCGCTCTGGT Chr2:119036260..119036279 59.92 55
upstream ENSMUSE00000562407 Chr2:119036517..119036567 TGACAATGCTTCCAAGTGGT Chr2:119036526..119036545 59.14 45
upstream ENSMUSE00000562406 Chr2:119036917..119037035 GCTTCGTCAGGAAAACAAGC Chr2:119037013..119037032 60 50

*** Putative Vector Insertion (Chr 2: 119037036 - 119037133) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000562405 Chr2:119037134..119037279 CTAGCCGTGCCTCTATCTCG Chr2:119037175..119037194 60.13 60
downstream ENSMUSE00000562403 Chr2:119037642..119037750 CTGGGTCCTGGTATCTGGTG Chr2:119037674..119037693 60.38 60
downstream ENSMUSE00000562401 Chr2:119040570..119040776 ACAGCCAATCTTTTGGCAAG Chr2:119040753..119040772 60.25 45
downstream ENSMUSE00000562400 Chr2:119041190..119041263 CTGCTGCATGACTCTCTGGA Chr2:119041266..119041285 60.29 55
downstream ENSMUSE00000562399 Chr2:119041945..119042043 TAAAGCCACTTGCCTCATCC Chr2:119041987..119042006 60.21 50
downstream ENSMUSE00000562398 Chr2:119042168..119042277 GCGCAAAGTAGCATCCTCAT Chr2:119042236..119042255 60.38 50
downstream ENSMUSE00000562397 Chr2:119042365..119042785 GCCATGACTCCTCCTCTCAC Chr2:119042651..119042670 59.8 60
downstream ENSMUSE00000685550 Chr2:119042365..119042785 GCCATGACTCCTCCTCTCAC Chr2:119042651..119042670 59.8 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCCTAGTGAGTGGGAGTGG Chr2:119037031..119037051 59.33 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGCCTAGTGAGTGGGAGTGG Chr2:119037031..119037051 59.33 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000068580