Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI13550
Trapped Gene
Chchd10 (ENSMUSG00000049422)
Vector Insertion
Chr 10: 75399191 - 75400051
Public Clones CMHD-GT_300C5-3 (cmhd) IST14743C6 (tigm)
Private Clones OST51971 (lexicon) OST28274 (lexicon) OST12120 (lexicon)
Severity of mutation (?) Insertion after 60% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000390495 (Chr10:75398983..75399190 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGTAGGGCATGTCATGGGTA Chr10:75399109..75399128 59.8 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000390495 (Chr10:75398983..75399190 +)
Downstram Exon
ENSMUSE00000350915 (Chr10:75400052..75400199 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGTAGGGCATGTCATGGGTA Chr10:75399109..75399128 59.8 50 GGTTAGGTCGCTCTGAGTGG Chr10:75400153..75400172 59.87 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000451333 Chr10:75398638..75398749 GCTACGGTCCACGCTTCTTA Chr10:75398660..75398679 60.41 55
upstream ENSMUSE00000390495 Chr10:75398983..75399190 TGTAGGGCATGTCATGGGTA Chr10:75399109..75399128 59.8 50

*** Putative Vector Insertion (Chr 10: 75399191 - 75400051) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000350915 Chr10:75400052..75400199 GGTTAGGTCGCTCTGAGTGG Chr10:75400153..75400172 59.87 60
downstream ENSMUSE00000451326 Chr10:75400320..75400489 CTCCCAGTCACATGGAACCT Chr10:75400419..75400438 59.96 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCAGTGGGGGAAATTCAGAG Chr10:75399148..75399168 60.04 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ATTCACACATGGTTGCATCG Chr10:75399224..75399244 60.4 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000049422