Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20908
Trapped Gene
Nipsnap1 (ENSMUSG00000034285)
Vector Insertion
Chr 11: 4783257 - 4783370
Public Clones (sanger)
Private Clones OST348536 (lexicon) OST287281 (lexicon) OST283241 (lexicon) OST280683 (lexicon)
OST260920 (lexicon) OST118084 (lexicon)
Severity of mutation (?) Insertion after 21% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000581370 (Chr11:4783222..4783256 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TTGCTTAGGAAGCTCCTGCT Chr11:4783230..4783249 59.36 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000581370 (Chr11:4783222..4783256 +)
Downstram Exon
ENSMUSE00000307937 (Chr11:4783371..4783498 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TTGCTTAGGAAGCTCCTGCT Chr11:4783230..4783249 59.36 50 TTGGACAGCAGAGTGGAGTG Chr11:4783469..4783488 60.02 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000406462 Chr11:4774006..4774106 GCTATTCACGAAGCCGAGAC Chr11:4774050..4774069 59.99 55
upstream ENSMUSE00000581372 Chr11:4782972..4783174 CGTGTCCATTCAGAGCAAAG Chr11:4783066..4783085 59.44 50
upstream ENSMUSE00000581370 Chr11:4783222..4783256 TTGCTTAGGAAGCTCCTGCT Chr11:4783230..4783249 59.36 50

*** Putative Vector Insertion (Chr 11: 4783257 - 4783370) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000307937 Chr11:4783371..4783498 TTGGACAGCAGAGTGGAGTG Chr11:4783469..4783488 60.02 55
downstream ENSMUSE00000307927 Chr11:4783729..4783774 TGTAGGCATCCAGACATTCG Chr11:4783767..4783786 59.67 50
downstream ENSMUSE00000307918 Chr11:4784025..4784119 CGTACCACGTGTTCCAGTTG Chr11:4784104..4784123 60.06 55
downstream ENSMUSE00000307909 Chr11:4788883..4788953 CTGAGAACCGCCATAGGTGT Chr11:4788906..4788925 60.13 55
downstream ENSMUSE00000715271 Chr11:4789100..4789240 ACTGGTTTCTCCTGGACAGC Chr11:4789160..4789179 59.31 55
downstream ENSMUSE00000718600 Chr11:4789100..4789240 ACTGGTTTCTCCTGGACAGC Chr11:4789160..4789179 59.31 55
downstream ENSMUSE00000656061 Chr11:4789536..4789567 CCCATTCAATCATGGTTCCT Chr11:4789560..4789579 59.6 45
downstream ENSMUSE00000656070 Chr11:4789536..4789567 CCCATTCAATCATGGTTCCT Chr11:4789560..4789579 59.6 45
downstream ENSMUSE00000656058 Chr11:4789895..4789989 TCTCCTGACGGTACTTGATGG Chr11:4789925..4789945 60.12 52.38
downstream ENSMUSE00000656069 Chr11:4789895..4789989 TCTCCTGACGGTACTTGATGG Chr11:4789925..4789945 60.12 52.38
downstream ENSMUSE00000656056 Chr11:4791406..4791489 TTCGAGTCTCCTCCCGAGAT Chr11:4791444..4791463 61.27 55
downstream ENSMUSE00000656068 Chr11:4791406..4791489 TTCGAGTCTCCTCCCGAGAT Chr11:4791444..4791463 61.27 55
downstream ENSMUSE00000307866 Chr11:4793123..4794203 TTGTCGAAGCTGAACACCAC Chr11:4793321..4793340 59.88 50
downstream ENSMUSE00000595105 Chr11:4793123..4794203 TTGTCGAAGCTGAACACCAC Chr11:4793321..4793340 59.88 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCTCCTGCTGTGAAATGTAGG Chr11:4783242..4783263 59.89 52.38 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCTCCTGCTGTGAAATGTAGG Chr11:4783242..4783263 59.89 52.38 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000034285