Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI2825
Trapped Gene
Glrx3 (ENSMUSG00000031068)
Vector Insertion
Chr 7: 144656191 - 144659615
Public Clones AE0543 (sanger) AG0355 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 95% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000205909 (Chr7:144656098..144656190 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTCAAATTGGCCAACCTACC Chr7:144656121..144656140 59.43 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000205909 (Chr7:144656098..144656190 +)
Downstram Exon
ENSMUSE00000587226 (Chr7:144659616..144659865 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTCAAATTGGCCAACCTACC Chr7:144656121..144656140 59.43 50 AGTCCTTGGACACCATGCTT Chr7:144659696..144659715 59.58 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000587230 Chr7:144629346..144629454 GAGCTACTGCGCCTCAAAAC Chr7:144629432..144629451 60.16 55
upstream ENSMUSE00000465947 Chr7:144636662..144636770 ATGAACGATGTCATGGCAGA Chr7:144636714..144636733 60.08 45
upstream ENSMUSE00000205914 Chr7:144644380..144644454 CCCACCTTCCTGTTCTTCAA Chr7:144644434..144644453 60.08 50
upstream ENSMUSE00000324353 Chr7:144645096..144645297 AGTCGACCGGTTAGATGGTG Chr7:144645107..144645126 59.99 55
upstream ENSMUSE00000587229 Chr7:144650806..144650978 CAGGGGCTCAAAACGTACTC Chr7:144650892..144650911 59.73 55
upstream ENSMUSE00000587228 Chr7:144651164..144651225 TCCCAAAGCACCCAAGTTAG Chr7:144651199..144651218 60.1 50
upstream ENSMUSE00000587227 Chr7:144653029..144653086 AAGCTTCCGTGATGCTCTTT Chr7:144653049..144653068 59.08 45
upstream ENSMUSE00000421474 Chr7:144654644..144654696 GAAGCAAAATGTGGATTCAGC Chr7:144654644..144654664 59.71 42.86
upstream ENSMUSE00000205918 Chr7:144655781..144655820 No primer for this exon
upstream ENSMUSE00000205909 Chr7:144656098..144656190 CTCAAATTGGCCAACCTACC Chr7:144656121..144656140 59.43 50

*** Putative Vector Insertion (Chr 7: 144656191 - 144659615) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000587226 Chr7:144659616..144659865 AGTCCTTGGACACCATGCTT Chr7:144659696..144659715 59.58 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGGGATCTTGTTGGAGGATT Chr7:144656159..144656179 60.13 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AAAGGTGACTCCGTGACTGG Chr7:144656231..144656251 60.15 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031068