Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24237
Trapped Gene
Btf3l4 (ENSMUSG00000028568)
Vector Insertion
Chr 4: 108491943 - 108498704
Public Clones (sanger)
Private Clones OST230279 (lexicon) OST203308 (lexicon) OST136551 (lexicon) OST122413 (lexicon)
OST122411 (lexicon) OST122297 (lexicon) OST78857 (lexicon) OST75019 (lexicon)
OST36108 (lexicon) OST21935 (lexicon)
Severity of mutation (?) Insertion after 35% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000525636 (Chr4:108498705..108498818 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACAGCTCGCAGGAAGAAGAA Chr4:108498796..108498815 60.28 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000525636 (Chr4:108498705..108498818 -)
Downstram Exon
ENSMUSE00000662464 (Chr4:108491741..108491942 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACAGCTCGCAGGAAGAAGAA Chr4:108498796..108498815 60.28 50 GCAAAGGTGTTAGCGGAGAG Chr4:108491838..108491857 60.01 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000662460 Chr4:108506113..108506159 No primer for this exon
upstream ENSMUSE00000662466 Chr4:108506113..108506219 GAGGCAGCCATCTTGCTCTT Chr4:108506150..108506169 61.97 55
upstream ENSMUSE00000662461 Chr4:108505748..108506169 CAGTGTCAGACGCTTTGAGC Chr4:108505789..108505808 59.78 55
upstream ENSMUSE00000662459 Chr4:108505018..108505056 No primer for this exon
upstream ENSMUSE00000662457 Chr4:108504311..108504536 TGCTGCACAGCTAAGATTGG Chr4:108504469..108504488 60.16 50
upstream ENSMUSE00000662465 Chr4:108503123..108503188 TCAAGCTCAGGTCCGGATAG Chr4:108503131..108503150 60.35 55
upstream ENSMUSE00000716151 Chr4:108503123..108503188 TCAAGCTCAGGTCCGGATAG Chr4:108503131..108503150 60.35 55
upstream ENSMUSE00000525636 Chr4:108498705..108498818 ACAGCTCGCAGGAAGAAGAA Chr4:108498796..108498815 60.28 50

*** Putative Vector Insertion (Chr 4: 108491943 - 108498704) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000662464 Chr4:108491741..108491942 GCAAAGGTGTTAGCGGAGAG Chr4:108491838..108491857 60.01 55
downstream ENSMUSE00000662463 Chr4:108490790..108490849 CTGGTTTGGGCGCTTTACTA Chr4:108490801..108490820 60.26 50
downstream ENSMUSE00000662456 Chr4:108489437..108489491 GCTTCATTTTTCGATGCTTCA Chr4:108489430..108489450 60.34 38.1
downstream ENSMUSE00000662458 Chr4:108489396..108489491 TCTAGTCCATGCCAGCTTCC Chr4:108489388..108489407 60.36 55
downstream ENSMUSE00000601484 Chr4:108489323..108489491 TCTAGTCCATGCCAGCTTCC Chr4:108489388..108489407 60.36 55
downstream ENSMUSE00000662462 Chr4:108489055..108489491 CAAATGACCGAGAACTGCTG Chr4:108489264..108489283 59.44 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGAACTAGCACACGCACAGG Chr4:108498654..108498674 59.11 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGAACTAGCACACGCACAGG Chr4:108498654..108498674 59.11 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 ACAGCTCGCAGGAAGAAGAA Chr4:108498794..108498814 60.28 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 GACAGCTACTGCTGCGTGAC Chr4:108498762..108498782 59.81 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028568