Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI7918
Trapped Gene
Mrps36 (ENSMUSG00000061474)
Vector Insertion
Chr 13: 101511187 - 101514484
Public Clones RRM130 (baygenomics) CMHD-GT_118A1-3 (cmhd) IST11813H7 (tigm) IST11813H7 (tigm)
Private Clones OST268858 (lexicon) OST144248 (lexicon) OST121335 (lexicon) OST121321 (lexicon)
OST121196 (lexicon) OST107013 (lexicon) OST58911 (lexicon) OST36337 (lexicon)
OST29884 (lexicon) OST29550 (lexicon) OST28833 (lexicon) OST21201 (lexicon)
OST18182 (lexicon) OST13658 (lexicon)
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000679942 (Chr13:101514485..101514612 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCTGCCCACTCCAGATACTT Chr13:101514571..101514590 59.31 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000679942 (Chr13:101514485..101514612 -)
Downstram Exon
ENSMUSE00000679953 (Chr13:101511123..101511186 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCTGCCCACTCCAGATACTT Chr13:101514571..101514590 59.31 55 TTAATGGAGCGTGTGGCTTC Chr13:101511140..101511159 61.17 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000679942 Chr13:101514485..101514612 GCTGCCCACTCCAGATACTT Chr13:101514571..101514590 59.31 55
upstream ENSMUSE00000679954 Chr13:101514485..101514614 GCTGCCCACTCCAGATACTT Chr13:101514571..101514590 59.31 55

*** Putative Vector Insertion (Chr 13: 101511187 - 101514484) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000679953 Chr13:101511123..101511186 TTAATGGAGCGTGTGGCTTC Chr13:101511140..101511159 61.17 50
downstream ENSMUSE00000461723 Chr13:101508969..101509156 TCAGTAAATCGGGGGACGTA Chr13:101509045..101509064 60.32 50
downstream ENSMUSE00000679941 Chr13:101508969..101509150 TCAGTAAATCGGGGGACGTA Chr13:101509045..101509064 60.32 50
downstream ENSMUSE00000679940 Chr13:101506032..101506228 GCTAACCCTTCTGTCTGGTGA Chr13:101506149..101506169 59.34 52.38
downstream ENSMUSE00000679946 Chr13:101505910..101506228 GCAGCTGACCGAGATACACA Chr13:101506009..101506028 60.02 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATGGTGCAGTAATCGCCTTG Chr13:101514422..101514442 61.05 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AACTTGTCAGGCTTGCTTGG Chr13:101514492..101514512 60.43 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GGGGTGGAGTTGTCCCTTAT Chr13:101514624..101514644 60.05 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 GGGGTGGAGTTGTCCCTTAT Chr13:101514624..101514644 60.05 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000061474