Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI29128
Trapped Gene
Dph3 (ENSMUSG00000021905)
Vector Insertion
Chr 14: 32896433 - 32897881
Public Clones IST14913B1 (tigm)
Private Clones OST7682 (lexicon)
Severity of mutation (?) Insertion after 63% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000690104 (Chr14:32897882..32898167 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGAAAAGGGCGGAAGAGTGT Chr14:32898005..32898024 60.25 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000690104 (Chr14:32897882..32898167 -)
Downstram Exon
ENSMUSE00000510619 (Chr14:32896355..32896432 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGAAAAGGGCGGAAGAGTGT Chr14:32898005..32898024 60.25 50 CCTTGTTGGTTGAAGGTGCT Chr14:32896362..32896381 60.15 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000121880 Chr14:32898596..32898795 GGGATAACTTTGCCATCACC Chr14:32898599..32898618 59.25 50
upstream ENSMUSE00000446599 Chr14:32898596..32898851 GGGATAACTTTGCCATCACC Chr14:32898599..32898618 59.25 50
upstream ENSMUSE00000517748 Chr14:32898093..32898167 CGTGTCCTAGCTGCTCACTC Chr14:32898117..32898136 58.78 60
upstream ENSMUSE00000690104 Chr14:32897882..32898167 AGAAAAGGGCGGAAGAGTGT Chr14:32898005..32898024 60.25 50

*** Putative Vector Insertion (Chr 14: 32896433 - 32897881) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000510619 Chr14:32896355..32896432 CCTTGTTGGTTGAAGGTGCT Chr14:32896362..32896381 60.15 50
downstream ENSMUSE00000383326 Chr14:32893752..32894455 TCTGGAAACGTCTGGGATTC Chr14:32894057..32894076 60.05 50
downstream ENSMUSE00000513766 Chr14:32893748..32896432 GGCAAGTTAGCAGGCTTCAC Chr14:32895418..32895437 60.02 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATAATCGCCTTGCAGCACAT Chr14:32897811..32897831 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CACTTTGGAAAGGGCAATAAA Chr14:32897862..32897883 59.11 38.1 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GCTCACTAATCGCCTTGCAG Chr14:32898103..32898123 61.07 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 ACGTGTCCTAGCTGCTCACC Chr14:32898116..32898136 60.48 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000021905