Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI5089
Trapped Gene
Zfp54 (ENSMUSG00000023882)
Vector Insertion
Chr 17: 21567286 - 21570366
Public Clones YTC632 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 9% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000554442 (Chr17:21567159..21567285 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAATGCCTGGACTCTGCTCA Chr17:21567210..21567229 61.52 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000554442 (Chr17:21567159..21567285 +)
Downstram Exon
ENSMUSE00000430298 (Chr17:21570367..21571967 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAATGCCTGGACTCTGCTCA Chr17:21567210..21567229 61.52 55 GGTTTGTTCCAGCACGAAGT Chr17:21570936..21570955 60.16 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000613179 Chr17:21560191..21560525 GCGTGATAGCTGGGACTGTT Chr17:21560247..21560266 60.28 55
upstream ENSMUSE00000322706 Chr17:21560346..21560414 TGATGCAGACCTGGAGTCAG Chr17:21560366..21560385 59.98 55
upstream ENSMUSE00000658150 Chr17:21560416..21560436 No primer for this exon
upstream ENSMUSE00000390836 Chr17:21560438..21560525 GGGAGGGCTGTAACCTACCA Chr17:21560505..21560524 61.26 60
upstream ENSMUSE00000554445 Chr17:21564701..21564792 No primer for this exon
upstream ENSMUSE00000613178 Chr17:21564701..21564792 No primer for this exon
upstream ENSMUSE00000554442 Chr17:21567159..21567285 GAATGCCTGGACTCTGCTCA Chr17:21567210..21567229 61.52 55

*** Putative Vector Insertion (Chr 17: 21567286 - 21570366) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000430298 Chr17:21570367..21571967 GGTTTGTTCCAGCACGAAGT Chr17:21570936..21570955 60.16 50
downstream ENSMUSE00000532895 Chr17:21570367..21572348 GGTTTGTTCCAGCACGAAGT Chr17:21570936..21570955 60.16 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AAGTAATCGCCTTGCAGCAC Chr17:21567334..21567354 60.42 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCATCGTGACTGGGAAAACC Chr17:21567333..21567353 60.9 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023882