Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI11784
Trapped Gene
Zfp652 (ENSMUSG00000075595)
Vector Insertion
Chr 11: 95614337 - 95614675
Public Clones A026E05 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 57% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000674206 (Chr11:95614189..95614336 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCATGGCTCATGTACGAAAA Chr11:95614307..95614326 59.54 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000674206 (Chr11:95614189..95614336 +)
Downstram Exon
ENSMUSE00000674205 (Chr11:95614676..95614791 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCATGGCTCATGTACGAAAA Chr11:95614307..95614326 59.54 45 GGAGTGCACCTTGAGTGACA Chr11:95614758..95614777 59.87 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000674202 Chr11:95573987..95574047 No primer for this exon
upstream ENSMUSE00000674201 Chr11:95610314..95611461 CCGTAGGAAAAGTGCAGAGC Chr11:95611197..95611216 60.01 55
upstream ENSMUSE00000268212 Chr11:95610381..95611461 CCGTAGGAAAAGTGCAGAGC Chr11:95611197..95611216 60.01 55
upstream ENSMUSE00000674206 Chr11:95614189..95614336 CCATGGCTCATGTACGAAAA Chr11:95614307..95614326 59.54 45

*** Putative Vector Insertion (Chr 11: 95614337 - 95614675) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000674205 Chr11:95614676..95614791 GGAGTGCACCTTGAGTGACA Chr11:95614758..95614777 59.87 55
downstream ENSMUSE00000674204 Chr11:95615225..95615369 CACTGGCACATGAACTGCTT Chr11:95615313..95615332 59.9 50
downstream ENSMUSE00000674200 Chr11:95624853..95633667 CTCCGCAGTGGAGTTCTTTC Chr11:95629170..95629189 59.99 55
downstream ENSMUSE00000674203 Chr11:95624853..95626025 CACCAAAAGCTGAACCGAAT Chr11:95625787..95625806 60.11 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGCTCTCTTAATCGCCTTGC Chr11:95614380..95614400 60.63 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTCTCCACCTGCAGACTCCT Chr11:95614356..95614376 59.99 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000075595