Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI10652
Trapped Gene
Eif2c1 (ENSMUSG00000041530)
Vector Insertion
Chr 4: 126141100 - 126145640
Public Clones (sanger) (sanger) (sanger) (sanger) (sanger) P009B11 (ggtc)
E056C08 (ggtc) (ggtc) E061D12 (ggtc) D138A11 (ggtc) (ggtc)
W143E07 (ggtc) E061D12 (ggtc) D067A08 (ggtc) (ggtc) (egtc)
IST10835G4 (tigm) IST14353G12 (tigm) IST14587E4 (tigm) IST10118B5 (tigm)
IST14469H12 (tigm) IST14117F11 (tigm) IST10986B9 (tigm) IST12033H7 (tigm)
IST13657D1 (tigm) IST14823A2 (tigm) IST10888C4 (tigm) IST14579G8 (tigm)
Private Clones OST422990 (lexicon) OST376097 (lexicon) OST333737 (lexicon) OST288054 (lexicon)
OST268886 (lexicon) OST240870 (lexicon) OST204316 (lexicon) OST119391 (lexicon)
OST113736 (lexicon)
Severity of mutation (?) Insertion after 1% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000368555 (Chr4:126145641..126145878 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTCACGCTGGACTCCTCAGT Chr4:126145701..126145720 60.61 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000368555 (Chr4:126145641..126145878 -)
Downstram Exon
ENSMUSE00000338399 (Chr4:126140916..126141099 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTCACGCTGGACTCCTCAGT Chr4:126145701..126145720 60.61 60 AGGACACTTATCCGGCTTGA Chr4:126140908..126140927 59.69 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

*** Putative Vector Insertion (Chr 4: 126141100 - 126145640) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000338399 Chr4:126140916..126141099 AGGACACTTATCCGGCTTGA Chr4:126140908..126140927 59.69 50
downstream ENSMUSE00000224186 Chr4:126138929..126139049 AGATCTGCGGTTTGAAATGC Chr4:126138984..126139003 60.22 45
downstream ENSMUSE00000224178 Chr4:126138150..126138331 ATAGATGCCAGGTGCCTCAT Chr4:126138131..126138150 59.54 50
downstream ENSMUSE00000224170 Chr4:126137611..126137747 CAGACTGGTGAAAGCCGAAC Chr4:126137628..126137647 60.83 55
downstream ENSMUSE00000342961 Chr4:126137245..126137379 TTGTAGAAGGCAGTGGCTGA Chr4:126137336..126137355 59.59 50
downstream ENSMUSE00000394898 Chr4:126137024..126137111 CACACGCGGTATTTCCTCTT Chr4:126137035..126137054 60.13 50
downstream ENSMUSE00000360501 Chr4:126136353..126136500 TCGAGGGGCAAATAGGTATG Chr4:126136332..126136351 59.92 50
downstream ENSMUSE00000228456 Chr4:126131549..126131668 CCCAGCCACAATGTTACAGA Chr4:126131626..126131645 59.57 50
downstream ENSMUSE00000372685 Chr4:126131070..126131192 CTCCGTCATGTCATCCTTCA Chr4:126131096..126131115 59.63 50
downstream ENSMUSE00000365915 Chr4:126130802..126130935 TGAGCACCTCTTCTCGACAC Chr4:126130781..126130800 59.13 55
downstream ENSMUSE00000341143 Chr4:126126011..126126195 AATCTTCCGAAGCTGGTCTG Chr4:126126146..126126165 59.43 50
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 TTTTAATCGCCTTGCAGCAC Chr4:126145572..126145592 61.26 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGGTCCTCTTTTTCGTGACTG Chr4:126145581..126145602 60.28 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 ATAATCGCCTTGCAGCACAT Chr4:126145809..126145829 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 TTAGGAGGCAAAAGGTGACG Chr4:126145826..126145846 60.24 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000041530