Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI19268
Trapped Gene
G430022H21Rik (ENSMUSG00000028114)
Vector Insertion
Chr 3: 123085761 - 123086513
Public Clones not available
Private Clones OST397977 (lexicon)
Severity of mutation (?) Insertion after 11% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000176349 (Chr3:123086514..123086602 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGAGAGTGCGGATAGCATTG Chr3:123086575..123086594 59.97 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000176349 (Chr3:123086514..123086602 -)
Downstram Exon
ENSMUSE00000176352 (Chr3:123085673..123085760 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGAGAGTGCGGATAGCATTG Chr3:123086575..123086594 59.97 50 CCGTTTTGAGTTTGGAGCAG Chr3:123085702..123085721 60.8 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000261154 Chr3:123088680..123088866 GTGAAAGTCTCCGGGTCAGA Chr3:123088847..123088866 60.24 55
upstream ENSMUSE00000176349 Chr3:123086514..123086602 TGAGAGTGCGGATAGCATTG Chr3:123086575..123086594 59.97 50

*** Putative Vector Insertion (Chr 3: 123085761 - 123086513) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000176352 Chr3:123085673..123085760 CCGTTTTGAGTTTGGAGCAG Chr3:123085702..123085721 60.8 50
downstream ENSMUSE00000176356 Chr3:123083566..123083646 TGCTGCATTTCCAGTTCATC Chr3:123083605..123083624 59.81 45
downstream ENSMUSE00000176364 Chr3:123083138..123083225 No primer for this exon
downstream ENSMUSE00000176347 Chr3:123079063..123079153 TGGGAGGAGTGTTTGACTTAGC Chr3:123079042..123079063 60.66 50
downstream ENSMUSE00000176346 Chr3:123077663..123077804 TTTCTCATTCGCAGTGATGC Chr3:123077659..123077678 59.96 45
downstream ENSMUSE00000176358 Chr3:123076888..123076980 TGCTGCAATCTCATCGATTT Chr3:123076926..123076945 59.38 40
downstream ENSMUSE00000176354 Chr3:123075475..123075591 CTCTGGAAAACTGCCTTTGG Chr3:123075460..123075479 59.85 50
downstream ENSMUSE00000176350 Chr3:123074193..123074403 CATGAATGAAGTCCCCGTCT Chr3:123074318..123074337 59.93 50
downstream ENSMUSE00000506094 Chr3:123071217..123072608 AGTATGAGTTGGGGGCACTG Chr3:123072500..123072519 59.99 55
downstream ENSMUSE00000563664 Chr3:123067864..123068061 TCCCTCTGAGGTCACATTCC Chr3:123067869..123067888 60.05 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GAGGGAGATTGCTGAAACCA Chr3:123086525..123086545 60.19 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAGGGAGATTGCTGAAACCA Chr3:123086525..123086545 60.19 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028114