Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14584
Trapped Gene
Rnf10 (ENSMUSG00000041740)
Vector Insertion
Chr 5: 115710127 - 115722327
Public Clones D060A05 (ggtc) D093C11 (ggtc) D093C11 (ggtc) FHCRC-GT-S8-11B1 (fhcrc)
Private Clones not available
Severity of mutation (?) Insertion after 22% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000650743 (Chr5:115722266..115722326 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGCGAGTCTAAACCCAAGAG Chr5:115722268..115722287 58.93 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000650743 (Chr5:115722266..115722326 -)
Downstram Exon
ENSMUSE00000268179 (Chr5:115710128..115710324 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGCGAGTCTAAACCCAAGAG Chr5:115722268..115722287 58.93 55 TTCACGCTTGCGATTGTAAC Chr5:115710256..115710275 59.88 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000690452 Chr5:115722355..115722380 No primer for this exon
upstream ENSMUSE00000650744 Chr5:115722328..115722903 GAAAAATGTCGCCATGAAGG Chr5:115722531..115722550 60.45 45
upstream ENSMUSE00000650743 Chr5:115722266..115722326 GGCGAGTCTAAACCCAAGAG Chr5:115722268..115722287 58.93 55
upstream ENSMUSE00000690456 Chr5:115722266..115722901 GAAAAATGTCGCCATGAAGG Chr5:115722531..115722550 60.45 45
upstream ENSMUSE00000690459 Chr5:115722266..115722907 GAAAAATGTCGCCATGAAGG Chr5:115722531..115722550 60.45 45
upstream ENSMUSE00000709339 Chr5:115722266..115722901 GAAAAATGTCGCCATGAAGG Chr5:115722531..115722550 60.45 45
upstream ENSMUSE00000268179 Chr5:115710128..115710324 TTACAATCGCAAGCGTGAAC Chr5:115710277..115710296 59.88 45
upstream ENSMUSE00000690450 Chr5:115710128..115710158 No primer for this exon

*** Putative Vector Insertion (Chr 5: 115710127 - 115722327) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000268172 Chr5:115706983..115707182 GCTCCTTGTTGAAAGGCTTG Chr5:115706977..115706996 59.99 50
downstream ENSMUSE00000268168 Chr5:115705359..115705449 TGTCAGGGTCAGCAAAGTGA Chr5:115705369..115705388 60.44 50
downstream ENSMUSE00000268162 Chr5:115701286..115701470 GTTATCTTGGCCGCAGTAGG Chr5:115701381..115701400 59.73 55
downstream ENSMUSE00000268151 Chr5:115701075..115701211 No primer for this exon
downstream ENSMUSE00000268142 Chr5:115699979..115700139 AGCAGCCTCAATGAAACAGG Chr5:115699972..115699991 60.4 50
downstream ENSMUSE00000268120 Chr5:115699406..115699531 AGCCACAACACCAGTGACCT Chr5:115699447..115699466 60.63 55
downstream ENSMUSE00000268111 Chr5:115698924..115699179 CTCCTCCTGGCTCATCTCAG Chr5:115698963..115698982 60.09 60
downstream ENSMUSE00000650742 Chr5:115698924..115699182 TTAGTGATGGGCTCCTCCTG Chr5:115698952..115698971 60.21 55
downstream ENSMUSE00000268104 Chr5:115698591..115698724 CAGACATGGAATACCCAGCA Chr5:115698571..115698590 59.52 50
downstream ENSMUSE00000268096 Chr5:115697178..115697295 TGCAGAGCCAGTTCACAGAT Chr5:115697197..115697216 59.58 50
downstream ENSMUSE00000268090 Chr5:115696889..115696990 CCATCTCAATTCTGCGCTCT Chr5:115696891..115696910 60.5 50
downstream ENSMUSE00000268085 Chr5:115695858..115696013 GGACCCAAAGCAGAATCAGA Chr5:115695904..115695923 60.19 50
downstream ENSMUSE00000268074 Chr5:115694030..115694130 TCGTCTTCCAGACTCCCAAC Chr5:115694033..115694052 60.24 55
downstream ENSMUSE00000268066 Chr5:115692643..115692700 No primer for this exon
downstream ENSMUSE00000453219 Chr5:115692402..115692560 AAGGCTGCTTCGATAGCTTG Chr5:115692421..115692440 59.75 50
downstream ENSMUSE00000495809 Chr5:115691786..115692070 GCCCCAAGCTAATCAATCTG Chr5:115691867..115691886 59.67 50
downstream ENSMUSE00000690447 Chr5:115691782..115692067 GCCCCAAGCTAATCAATCTG Chr5:115691867..115691886 59.67 50
downstream ENSMUSE00000690454 Chr5:115691779..115692067 GCCCCAAGCTAATCAATCTG Chr5:115691867..115691886 59.67 50
downstream ENSMUSE00000690458 Chr5:115691421..115692070 TGTAAAGAGGGGAAGCTGGA Chr5:115691786..115691805 59.81 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTTACAATCGCAAGCGTGAA Chr5:115719276..115719296 59.88 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGAGAACGTGACTGGGAAAAC Chr5:115719261..115719282 60.14 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000041740