Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI34919
Trapped Gene
Eif2c1 (ENSMUSG00000041530)
Vector Insertion
Chr 4: 126126010 - 126130936
Public Clones IST14472H4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 54% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000365915 (Chr4:126130802..126130935 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGTCGAGAAGAGGTGCTCA Chr4:126130803..126130822 59.13 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000365915 (Chr4:126130802..126130935 -)
Downstram Exon
ENSMUSE00000341143 (Chr4:126126011..126126195 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGTCGAGAAGAGGTGCTCA Chr4:126130803..126130822 59.13 55 AATCTTCCGAAGCTGGTCTG Chr4:126126146..126126165 59.43 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000368555 Chr4:126145641..126145878 CTCACGCTGGACTCCTCAGT Chr4:126145701..126145720 60.61 60
upstream ENSMUSE00000338399 Chr4:126140916..126141099 TCAAGCCGGATAAGTGTCCT Chr4:126140930..126140949 59.69 50
upstream ENSMUSE00000224186 Chr4:126138929..126139049 GATCGCAAGCCTGTGTATGA Chr4:126138981..126139000 59.83 50
upstream ENSMUSE00000224178 Chr4:126138150..126138331 ATGAGGCACCTGGCATCTAT Chr4:126138153..126138172 59.54 50
upstream ENSMUSE00000224170 Chr4:126137611..126137747 TTCGGCTTTCACCAGTCTGT Chr4:126137649..126137668 60.83 50
upstream ENSMUSE00000342961 Chr4:126137245..126137379 AGCGTGTTCGGTTTACCAAG Chr4:126137255..126137274 60.17 50
upstream ENSMUSE00000394898 Chr4:126137024..126137111 AAGAGGAAATACCGCGTGTG Chr4:126137057..126137076 60.13 50
upstream ENSMUSE00000360501 Chr4:126136353..126136500 GCCTACAAGTTGGGCAAGAA Chr4:126136380..126136399 60.25 50
upstream ENSMUSE00000228456 Chr4:126131549..126131668 AGCGGTGCATTAAGAAGCTG Chr4:126131627..126131646 60.54 50
upstream ENSMUSE00000372685 Chr4:126131070..126131192 ATGACATGACGGAGGTGACA Chr4:126131112..126131131 59.96 50
upstream ENSMUSE00000365915 Chr4:126130802..126130935 GTGTCGAGAAGAGGTGCTCA Chr4:126130803..126130822 59.13 55
upstream ENSMUSE00000341143 Chr4:126126011..126126195 CAGCCATGCTTCTGCAAATA Chr4:126126118..126126137 59.98 45

*** Putative Vector Insertion (Chr 4: 126126010 - 126130936) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000369122 Chr4:126120325..126120484 CTCCGACACGTTTCACTTCA Chr4:126120442..126120461 59.87 50
downstream ENSMUSE00000234783 Chr4:126120060..126120150 TCTGCTCCCAGGAAAATCAC Chr4:126120087..126120106 60.19 50
downstream ENSMUSE00000234774 Chr4:126119133..126119327 GAAGCGGGTGGACTTGTAGA Chr4:126119168..126119187 60.26 55
downstream ENSMUSE00000234766 Chr4:126118421..126118555 GGCAAGCAGCTCATAGTGAA Chr4:126118510..126118529 59.18 50
downstream ENSMUSE00000377612 Chr4:126117608..126117709 GGTGATGTTGGTGTCCACAG Chr4:126117634..126117653 59.85 55
downstream ENSMUSE00000524941 Chr4:126117086..126117285 AAACGGTTGTCATCCCAAAG Chr4:126117214..126117233 59.83 45
downstream ENSMUSE00000631261 Chr4:126112256..126116855 GTGTTACGAGGGCTGGATGT Chr4:126116060..126116079 60 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCCCTCTGCCTATCCCTAGA Chr4:126127933..126127953 60.55 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCCCTCTGCCTATCCCTAGA Chr4:126127933..126127953 60.55 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

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