Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI36257
Trapped Gene
Morn3 (ENSMUSG00000029477)
Vector Insertion
Chr 5: 123489265 - 123491250
Public Clones IST14500C2 (tigm) IST14500D2 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 42% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000649828 (Chr5:123491092..123491249 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGAAGCGAGATGGTTATGGA Chr5:123491170..123491189 60.04 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000649828 (Chr5:123491092..123491249 -)
Downstram Exon
ENSMUSE00000189240 (Chr5:123489266..123489425 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGAAGCGAGATGGTTATGGA Chr5:123491170..123491189 60.04 50 CCGAAAAACTGGATCCCATA Chr5:123489381..123489400 59.76 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000497370 Chr5:123496640..123496829 AAAGCCCAGAAGAACGGACT Chr5:123496708..123496727 60.25 50
upstream ENSMUSE00000649828 Chr5:123491092..123491249 GGAAGCGAGATGGTTATGGA Chr5:123491170..123491189 60.04 50
upstream ENSMUSE00000189240 Chr5:123489266..123489425 GTTTTTCGGACCCAAGGAGT Chr5:123489392..123489411 60.34 50

*** Putative Vector Insertion (Chr 5: 123489265 - 123491250) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000328409 Chr5:123487685..123487869 GTCCACCCAGTAGCCTTCAA Chr5:123487741..123487760 60.11 55
downstream ENSMUSE00000495618 Chr5:123487142..123487369 AGATTTGCCCTCTCTGGTCA Chr5:123487181..123487200 59.8 50
downstream ENSMUSE00000537470 Chr5:123487142..123487369 AGATTTGCCCTCTCTGGTCA Chr5:123487181..123487200 59.8 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGTGGTGCGGTCTATGAAGG Chr5:123491202..123491222 60.13 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGTGGTGCGGTCTATGAAGG Chr5:123491202..123491222 60.13 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029477