Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31950
Trapped Gene
Adamts14 (ENSMUSG00000059901)
Vector Insertion
Chr 10: 60663923 - 60665779
Public Clones IST15088E11 (tigm) IST10224E3 (tigm) IST14320A4 (tigm) IST11732C12 (tigm)
IST14320A4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 80% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000500118 (Chr10:60665571..60665778 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAAGAGTGGGGTGCCTGTAG Chr10:60665749..60665768 59.72 60 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000500118 (Chr10:60665571..60665778 -)
Downstram Exon
ENSMUSE00000505959 (Chr10:60663924..60664053 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAAGAGTGGGGTGCCTGTAG Chr10:60665749..60665768 59.72 60 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000666275 Chr10:60735904..60736186 No primer for this exon
upstream ENSMUSE00000713901 Chr10:60735904..60735990 No primer for this exon
upstream ENSMUSE00000576050 Chr10:60733605..60734005 AGATTTCCAGGGACGCTTCT Chr10:60733931..60733950 60.21 50
upstream ENSMUSE00000576049 Chr10:60712352..60712508 TGGTGACCTTCACAACGAAG Chr10:60712352..60712371 59.72 50
upstream ENSMUSE00000576048 Chr10:60708674..60708864 CAGGGAGCATACGCAGAACT Chr10:60708697..60708716 60.42 55
upstream ENSMUSE00000482142 Chr10:60693166..60693249 No primer for this exon
upstream ENSMUSE00000576042 Chr10:60692404..60692551 CTGAACATCACGACCACGTC Chr10:60692438..60692457 60.16 55
upstream ENSMUSE00000717226 Chr10:60692395..60692551 CTGAACATCACGACCACGTC Chr10:60692438..60692457 60.16 55
upstream ENSMUSE00000576047 Chr10:60689855..60689960 GCCACCCACTAAGAAGCTGT Chr10:60689917..60689936 59.36 55
upstream ENSMUSE00000491823 Chr10:60688085..60688228 CCACAGGTTCCACTGGTCTC Chr10:60688119..60688138 60.56 60
upstream ENSMUSE00000490386 Chr10:60687021..60687153 ACGATTGCCTCCTTGATGAC Chr10:60687129..60687148 60.08 50
upstream ENSMUSE00000576046 Chr10:60685532..60685645 CCCATACTTCTGCAAGACCAA Chr10:60685575..60685595 60.12 47.62
upstream ENSMUSE00000486699 Chr10:60684606..60684754 CTCATGTTCTCGGTCGTGTG Chr10:60684649..60684668 60.31 55
upstream ENSMUSE00000484980 Chr10:60681106..60681281 TGCCAAGCGAAACTCCTACT Chr10:60681157..60681176 60.01 50
upstream ENSMUSE00000507297 Chr10:60676278..60676411 AGTCTGCCGACACTGGAGAT Chr10:60676365..60676384 59.87 55
upstream ENSMUSE00000513248 Chr10:60675872..60675995 TGAAGACGGATGACAAGTGC Chr10:60675945..60675964 59.84 50
upstream ENSMUSE00000512229 Chr10:60674336..60674416 CTCTCAAACAGGTGCAGATCC Chr10:60674393..60674413 59.86 52.38
upstream ENSMUSE00000518023 Chr10:60670048..60670211 CCTAGAGTGGGAGCACGAAG Chr10:60670110..60670129 60.01 60
upstream ENSMUSE00000509589 Chr10:60668112..60668280 GAGGGTAAACCCCGAGGTAG Chr10:60668246..60668265 59.82 60
upstream ENSMUSE00000508548 Chr10:60667595..60667727 CCATAAAAAGAGGCCCAAGC Chr10:60667638..60667657 60.92 50
upstream ENSMUSE00000500118 Chr10:60665571..60665778 GAAGAGTGGGGTGCCTGTAG Chr10:60665749..60665768 59.72 60
upstream ENSMUSE00000505959 Chr10:60663924..60664053 GGTGGTATGCAGGAACACTTC Chr10:60663995..60664015 59.47 52.38

*** Putative Vector Insertion (Chr 10: 60663923 - 60665779) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000511920 Chr10:60663111..60663221 ACCTCCGCCTTCACTGTAGA Chr10:60663166..60663185 59.87 55
downstream ENSMUSE00000712810 Chr10:60661216..60661707 ACGGTCCAACACTTCCATTC Chr10:60661630..60661649 59.83 50
downstream ENSMUSE00000576036 Chr10:60659860..60661707 CTGGTCTAACCAGGGACCAA Chr10:60660411..60660430 59.96 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGAAGAGTGGGGTGCCTGTA Chr10:60665748..60665768 59.72 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGAAGAGTGGGGTGCCTGTA Chr10:60665748..60665768 59.72 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000059901