Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31072
Trapped Gene
Rps10 (ENSMUSG00000052146)
Vector Insertion
Chr 17: 27772148 - 27773575
Public Clones IST12359E11 (tigm) IST12600F12 (tigm) IST13611H1 (tigm) IST12139A1 (tigm)
IST13738A1 (tigm) IST10927B10 (tigm) IST13788C10 (tigm) IST13688A9 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000700896 (Chr17:27773510..27773574 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAGTCCCCCATCAGACAACT Chr17:27773521..27773540 59.96 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000700896 (Chr17:27773510..27773574 -)
Downstram Exon
ENSMUSE00000700904 (Chr17:27772149..27772221 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAGTCCCCCATCAGACAACT Chr17:27773521..27773540 59.96 55 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000700896 Chr17:27773510..27773574 CAGTCCCCCATCAGACAACT Chr17:27773521..27773540 59.96 55
upstream ENSMUSE00000700904 Chr17:27772149..27772221 No primer for this exon
upstream ENSMUSE00000700926 Chr17:27772149..27772185 No primer for this exon

*** Putative Vector Insertion (Chr 17: 27772148 - 27773575) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000549878 Chr17:27771271..27771420 CATCACGCCCTCCTTAAAAA Chr17:27771336..27771355 60.07 45
downstream ENSMUSE00000721935 Chr17:27771271..27771420 CATCACGCCCTCCTTAAAAA Chr17:27771336..27771355 60.07 45
downstream ENSMUSE00000139792 Chr17:27771017..27771188 ATGCCCTCGTTCGTAAGGTA Chr17:27771090..27771109 59.59 50
downstream ENSMUSE00000139788 Chr17:27769224..27769301 GCGCTCCTTCTGTAGGTGTC Chr17:27769210..27769229 60.02 60
downstream ENSMUSE00000700902 Chr17:27768145..27768200 GCCTCAGCTTTCTTGTCAGC Chr17:27768154..27768173 60.28 55
downstream ENSMUSE00000139784 Chr17:27767374..27767451 AACTTCACTGAGGTGGCTGA Chr17:27767384..27767403 58.44 50
downstream ENSMUSE00000700895 Chr17:27767369..27767451 AACTTCACTGAGGTGGCTGA Chr17:27767384..27767403 58.44 50
downstream ENSMUSE00000700898 Chr17:27767367..27767451 AACTTCACTGAGGTGGCTGA Chr17:27767384..27767403 58.44 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATAATCGCCTTGCAGCACAT Chr17:27773505..27773525 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ATCTCAGTCCCCCATCAGAC Chr17:27773523..27773543 58.9 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000052146