Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6211
Trapped Gene
Btf3l4 (ENSMUSG00000028568)
Vector Insertion
Chr 4: 108506112 - 108506170
Public Clones CSG280 (baygenomics) H003A03 (ggtc) Q012E01 (ggtc) P147G04 (ggtc)
P027A10 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000662466 (Chr4:108506113..108506219 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGGCAGCCATCTTGCTCTT Chr4:108506150..108506169 61.97 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000662466 (Chr4:108506113..108506219 -)
Downstram Exon
ENSMUSE00000662461 (Chr4:108505748..108506169 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGGCAGCCATCTTGCTCTT Chr4:108506150..108506169 61.97 55 ATCTGCTGCTCAAAGCGTCT Chr4:108505760..108505779 60.31 50

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

*** Putative Vector Insertion (Chr 4: 108506112 - 108506170) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000662461 Chr4:108505748..108506169 ATCTGCTGCTCAAAGCGTCT Chr4:108505760..108505779 60.31 50
downstream ENSMUSE00000662459 Chr4:108505018..108505056 No primer for this exon
downstream ENSMUSE00000662457 Chr4:108504311..108504536 TGTGCCAATCTTAGCTGTGC Chr4:108504443..108504462 60.02 50
downstream ENSMUSE00000662465 Chr4:108503123..108503188 CTATCCGGACCTGAGCTTGA Chr4:108503109..108503128 60.35 55
downstream ENSMUSE00000716151 Chr4:108503123..108503188 CTATCCGGACCTGAGCTTGA Chr4:108503109..108503128 60.35 55
downstream ENSMUSE00000525636 Chr4:108498705..108498818 TTCTTCTTCCTGCGAGCTGT Chr4:108498774..108498793 60.28 50
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 GAGGCAGCCATCTTGCTCTT Chr4:108506148..108506168 61.97 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAGGCAGCCATCTTGCTCTT Chr4:108506148..108506168 61.97 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028568