Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI36344
Trapped Gene
Klhl12 (ENSMUSG00000026455)
Vector Insertion
Chr 1: 136372480 - 136380323
Public Clones IST13435C1 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 49% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000159327 (Chr1:136372365..136372479 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGATGCAAGGACCCAGAACA Chr1:136372447..136372466 60.66 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000159327 (Chr1:136372365..136372479 +)
Downstram Exon
ENSMUSE00000159340 (Chr1:136380324..136380430 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGATGCAAGGACCCAGAACA Chr1:136372447..136372466 60.66 50 CTCCTGTGTCTTGGGGTCAT Chr1:136380418..136380437 59.96 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000539138 Chr1:136352108..136352242 TGGAGTTTACCACGGAGACTG Chr1:136352120..136352140 60.16 52.38
upstream ENSMUSE00000706808 Chr1:136352132..136352242 No primer for this exon
upstream ENSMUSE00000690839 Chr1:136355005..136355243 GCAACACCCTCTGTGATGTG Chr1:136355134..136355153 60.16 55
upstream ENSMUSE00000711567 Chr1:136355005..136355243 GCAACACCCTCTGTGATGTG Chr1:136355134..136355153 60.16 55
upstream ENSMUSE00000159332 Chr1:136355043..136355243 GCAACACCCTCTGTGATGTG Chr1:136355134..136355153 60.16 55
upstream ENSMUSE00000159329 Chr1:136360404..136360557 AGATCCTGCTGGACTTCGTG Chr1:136360462..136360481 60.41 55
upstream ENSMUSE00000159333 Chr1:136364228..136364445 TCAAGTGCGACGAGATTCAG Chr1:136364426..136364445 60.14 50
upstream ENSMUSE00000159339 Chr1:136368971..136369120 GCTGACCCCCAGGTACATTA Chr1:136369081..136369100 59.81 55
upstream ENSMUSE00000159327 Chr1:136372365..136372479 AGATGCAAGGACCCAGAACA Chr1:136372447..136372466 60.66 50

*** Putative Vector Insertion (Chr 1: 136372480 - 136380323) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000159340 Chr1:136380324..136380430 CTCCTGTGTCTTGGGGTCAT Chr1:136380418..136380437 59.96 55
downstream ENSMUSE00000159324 Chr1:136382295..136382490 AGGCCACATACCGTCTCTTG Chr1:136382328..136382347 60.13 55
downstream ENSMUSE00000159335 Chr1:136383587..136383745 GATTATTCCGCTGGCCACTA Chr1:136383738..136383757 60.06 50
downstream ENSMUSE00000159338 Chr1:136384221..136384319 No primer for this exon
downstream ENSMUSE00000379981 Chr1:136385512..136385698 GAGTCTCCCTCGAAGCACTG Chr1:136385687..136385706 60.13 60
downstream ENSMUSE00000690837 Chr1:136385593..136385698 GAGTCTCCCTCGAAGCACTG Chr1:136385687..136385706 60.13 60
downstream ENSMUSE00000331773 Chr1:136385909..136387595 CGAAGGAGAGTGTGCATCAA Chr1:136386203..136386222 59.98 50
downstream ENSMUSE00000690835 Chr1:136385909..136387450 CGAAGGAGAGTGTGCATCAA Chr1:136386203..136386222 59.98 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGATGCAAGGACCCAGAACA Chr1:136378448..136378468 60.66 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCTTTTTCGTGACTGGGAAA Chr1:136378524..136378544 60.08 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026455