Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI29510
Trapped Gene
Dnajb6 (ENSMUSG00000029131)
Vector Insertion
Chr 5: 30093644 - 30107669
Public Clones 5SP124H05 (ggtc) P124G05 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 100% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000700759 (Chr5:30092865..30093643 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTAAGGAGCACCTGCTACGC Chr5:30092936..30092955 60.04 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000700759 (Chr5:30092865..30093643 +)
Downstram Exon
ENSMUSE00000655109 (Chr5:30107670..30107990 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTAAGGAGCACCTGCTACGC Chr5:30092936..30092955 60.04 60 CCTGTCCTGCTCGTCATCTT Chr5:30107950..30107969 60.41 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000700760 Chr5:30062480..30062656 TGACGCACTTCCTGTTTGTT Chr5:30062508..30062527 59.34 45
upstream ENSMUSE00000185970 Chr5:30062568..30062656 No primer for this exon
upstream ENSMUSE00000185974 Chr5:30074885..30074975 TGCCTCACCTGAGGACATTA Chr5:30074949..30074968 59.24 50
upstream ENSMUSE00000710480 Chr5:30074885..30074975 TGCCTCACCTGAGGACATTA Chr5:30074949..30074968 59.24 50
upstream ENSMUSE00000185969 Chr5:30077478..30077587 AAGCAGAGCGGAAGTTCAAA Chr5:30077534..30077553 60.13 45
upstream ENSMUSE00000185979 Chr5:30078937..30078999 GCGGGACATCTACGACAAAT Chr5:30078944..30078963 59.96 50
upstream ENSMUSE00000185975 Chr5:30080082..30080192 CGGAACCCAGATGATGTCTT Chr5:30080126..30080145 59.93 50
upstream ENSMUSE00000185978 Chr5:30082774..30082878 GGATTTTCGCACAGGAAAAG Chr5:30082807..30082826 59.69 45
upstream ENSMUSE00000185980 Chr5:30084088..30084419 GCTCAGCGAGGAGTATGAGG Chr5:30084338..30084357 60.12 60
upstream ENSMUSE00000185972 Chr5:30090939..30091070 GGTCCCCGAGGAAATAGAAG Chr5:30090974..30090993 59.9 55
upstream ENSMUSE00000185973 Chr5:30092046..30092187 CATGGGGGTCTCACTTCATT Chr5:30092072..30092091 59.78 50
upstream ENSMUSE00000655112 Chr5:30092865..30092935 TGTGGAGAACGGTCAAGAAA Chr5:30092868..30092887 59.26 45
upstream ENSMUSE00000700759 Chr5:30092865..30093643 GTAAGGAGCACCTGCTACGC Chr5:30092936..30092955 60.04 60

*** Putative Vector Insertion (Chr 5: 30093644 - 30107669) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000655109 Chr5:30107670..30107990 CCTGTCCTGCTCGTCATCTT Chr5:30107950..30107969 60.41 55
downstream ENSMUSE00000655108 Chr5:30111571..30111653 TCCTCTTGCCACCTTCTTTG Chr5:30111597..30111616 60.37 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGACACGTTAGGGCATACCA Chr5:30093629..30093649 59.99 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGACACGTTAGGGCATACCA Chr5:30093629..30093649 59.99 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029131