Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI29306
Trapped Gene
Magix (ENSMUSG00000031147)
Vector Insertion
Chr X: 7252318 - 7252966
Public Clones CMHD-GT_424C3-3 (cmhd) PSTVUpb20d4 (vanderbilt) IST13663C1 (tigm) IST14564G3 (tigm)
IST12230F9 (tigm) IST14445G11 (tigm) IST14154H5 (tigm) IST14564G3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 71% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000206875 (ChrX:7252967..7253135 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGGAGAGGATTCGAACTGG ChrX:7253053..7253072 59.65 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000206875 (ChrX:7252967..7253135 -)
Downstram Exon
ENSMUSE00000394744 (ChrX:7251754..7252317 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGGAGAGGATTCGAACTGG ChrX:7253053..7253072 59.65 55 GGGTACCAGGGATACGGTTT ChrX:7252116..7252135 59.94 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000703884 ChrX:7258272..7258350 GACTTTACAGCTCCCGCAAC ChrX:7258295..7258314 59.88 55
upstream ENSMUSE00000206874 ChrX:7258076..7258215 CCCCTGCGCTAAAAACTAATC ChrX:7258163..7258183 60.1 47.62
upstream ENSMUSE00000206873 ChrX:7257026..7257172 GACAGCGCTAGTACGAAAAGC ChrX:7257058..7257078 59.33 52.38
upstream ENSMUSE00000206872 ChrX:7254246..7254328 GTCGCTTGCCTCATGCTACT ChrX:7254267..7254286 60.57 55
upstream ENSMUSE00000206876 ChrX:7253226..7253462 TTTAAGTGGAGGCCGAAATG ChrX:7253300..7253319 60.07 45
upstream ENSMUSE00000206875 ChrX:7252967..7253135 GTGGAGAGGATTCGAACTGG ChrX:7253053..7253072 59.65 55

*** Putative Vector Insertion (Chr X: 7252318 - 7252966) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000394744 ChrX:7251754..7252317 GGGTACCAGGGATACGGTTT ChrX:7252116..7252135 59.94 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAGAGAGGTAATCGCCTTGC ChrX:7252903..7252923 59.98 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CATCGGAAGACAGGTGAGGT ChrX:7252958..7252978 60.11 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GCCTTATAATCGCCTTGCAG ChrX:7253071..7253091 59.84 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 AACACAGGGCCTTACGTGAC ChrX:7253079..7253099 60.03 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031147