Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI7064
Trapped Gene
Mphosph6 (ENSMUSG00000031843)
Vector Insertion
Chr 8: 120318572 - 120322943
Public Clones RRZ074 (baygenomics)
Private Clones OST59329 (lexicon)
Severity of mutation (?) Insertion after 34% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000320725 (Chr8:120322944..120323056 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGGACTCGGAAACCAAGAA Chr8:120323022..120323041 60.22 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000320725 (Chr8:120322944..120323056 -)
Downstram Exon
ENSMUSE00000320718 (Chr8:120318481..120318571 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGGACTCGGAAACCAAGAA Chr8:120323022..120323041 60.22 50 TCAGGATTAAACCCCCTGAA Chr8:120318466..120318485 59.36 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000320735 Chr8:120325752..120325829 TCGGAGCGCAAGACTAAGTT Chr8:120325777..120325796 60.15 50
upstream ENSMUSE00000320725 Chr8:120322944..120323056 CTGGACTCGGAAACCAAGAA Chr8:120323022..120323041 60.22 50

*** Putative Vector Insertion (Chr 8: 120318572 - 120322943) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000320718 Chr8:120318481..120318571 TCAGGATTAAACCCCCTGAA Chr8:120318466..120318485 59.36 45
downstream ENSMUSE00000213654 Chr8:120316579..120316676 TCGACCTCTACCGTCTCGTC Chr8:120316584..120316603 60.41 60
downstream ENSMUSE00000320701 Chr8:120315545..120316218 ACCAGTACCGACGACAGGAG Chr8:120315988..120316007 60.17 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CGAGCACTGGTACCTGGATT Chr8:120322964..120322984 60.13 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CGAGCACTGGTACCTGGATT Chr8:120322964..120322984 60.13 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031843