Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI9962
Trapped Gene
Glrx3 (ENSMUSG00000031068)
Vector Insertion
Chr 7: 144645298 - 144650805
Public Clones XC486 (baygenomics) XC436 (baygenomics) PST12529-NL (escells)
Private Clones not available
Severity of mutation (?) Insertion after 48% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000324353 (Chr7:144645096..144645297 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGTCGACCGGTTAGATGGTG Chr7:144645107..144645126 59.99 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000324353 (Chr7:144645096..144645297 +)
Downstram Exon
ENSMUSE00000587229 (Chr7:144650806..144650978 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGTCGACCGGTTAGATGGTG Chr7:144645107..144645126 59.99 55 GTACGTTTTGAGCCCCTGTC Chr7:144650912..144650931 59.6 55

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

*** Putative Vector Insertion (Chr 7: 144645298 - 144650805) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000587229 Chr7:144650806..144650978 GTACGTTTTGAGCCCCTGTC Chr7:144650912..144650931 59.6 55
downstream ENSMUSE00000587228 Chr7:144651164..144651225 CCAGCTCTTCTGATGCTTCC Chr7:144651191..144651210 60.1 55
downstream ENSMUSE00000587227 Chr7:144653029..144653086 CGGAAGCTTTATTGGTCAGC Chr7:144653060..144653079 59.85 50
downstream ENSMUSE00000421474 Chr7:144654644..144654696 TGCTGAATCCACATTTTGCT Chr7:144654668..144654687 59.28 40
downstream ENSMUSE00000205918 Chr7:144655781..144655820 No primer for this exon
downstream ENSMUSE00000205909 Chr7:144656098..144656190 GGTAGGTTGGCCAATTTGAG Chr7:144656143..144656162 59.43 50
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 TAATCGCCTTGCAGCACATC Chr7:144645349..144645369 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector

Come back to gene ENSMUSG00000031068