Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI30320
Trapped Gene
Klf3 (ENSMUSG00000029178)
Vector Insertion
Chr 5: 65214269 - 65218348
Public Clones D014C10 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 67% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000186619 (Chr5:65214118..65214268 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCCCCTTTAATGAACCCAGT Chr5:65214222..65214241 60.05 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000186619 (Chr5:65214118..65214268 +)
Downstram Exon
ENSMUSE00000186625 (Chr5:65218349..65218509 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCCCCTTTAATGAACCCAGT Chr5:65214222..65214241 60.05 50 CTGTGTATCCTGCGCTTCCT Chr5:65218438..65218457 60.42 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000394739 Chr5:65194628..65195030 GTCATGTGACTGCCCAGAGTT Chr5:65194780..65194800 60.17 52.38
upstream ENSMUSE00000186617 Chr5:65207974..65208069 ATCCAGTCCCTGTCAAGCAG Chr5:65208026..65208045 60.26 55
upstream ENSMUSE00000186613 Chr5:65213113..65213596 GTCCAGCCCGTTCCTTTTAT Chr5:65213500..65213519 60.32 50
upstream ENSMUSE00000186619 Chr5:65214118..65214268 CCCCCTTTAATGAACCCAGT Chr5:65214222..65214241 60.05 50

*** Putative Vector Insertion (Chr 5: 65214269 - 65218348) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000186625 Chr5:65218349..65218509 CTGTGTATCCTGCGCTTCCT Chr5:65218438..65218457 60.42 55
downstream ENSMUSE00000186622 Chr5:65220216..65221367 ACCCTCTCGGTATCCAGCTT Chr5:65221268..65221287 60.1 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTAATCGCCTTGCAGCACAT Chr5:65214319..65214339 61.2 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCTTATTCTCCATTGCGTGAC Chr5:65214305..65214326 59.58 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029178