Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI10546
Trapped Gene
Creld1 (ENSMUSG00000030284)
Vector Insertion
Chr 6: 113434572 - 113438064
Public Clones M090B02 (ggtc) G046E05 (ggtc) G077G08 (ggtc) G046B01 (ggtc) IST14872H5 (tigm)
IST10316G2 (tigm) IST12094C4BBF1 (tigm) IST10055B6 (tigm) IST12094F8 (tigm)
IST13438C4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 20% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000197059 (Chr6:113434489..113434571 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGAACCATCCGGGACAACTT Chr6:113434498..113434517 60.75 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000197059 (Chr6:113434489..113434571 +)
Downstram Exon
ENSMUSE00000197055 (Chr6:113438065..113438175 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGAACCATCCGGGACAACTT Chr6:113434498..113434517 60.75 50 AACCACCAGTTTTCCACCAG Chr6:113438172..113438191 59.86 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000224405 Chr6:113433554..113434070 GCACTGGTGGACAACTTCAA Chr6:113434047..113434066 59.73 50
upstream ENSMUSE00000197059 Chr6:113434489..113434571 AGAACCATCCGGGACAACTT Chr6:113434498..113434517 60.75 50

*** Putative Vector Insertion (Chr 6: 113434572 - 113438064) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000197055 Chr6:113438065..113438175 AACCACCAGTTTTCCACCAG Chr6:113438172..113438191 59.86 50
downstream ENSMUSE00000197051 Chr6:113438473..113438564 CTTCAGGGAATCGGAACAGA Chr6:113438527..113438546 60.19 50
downstream ENSMUSE00000197061 Chr6:113439428..113439604 AAGTAGCCAAGGCCACACTG Chr6:113439569..113439588 60.31 55
downstream ENSMUSE00000197065 Chr6:113439689..113439784 CACTGCAGACAATGGGATTC Chr6:113439749..113439768 59.09 50
downstream ENSMUSE00000197063 Chr6:113441647..113441730 GTGGCTTGCTCTGTACCACA Chr6:113441680..113441699 59.9 55
downstream ENSMUSE00000197052 Chr6:113441888..113441983 No primer for this exon
downstream ENSMUSE00000197050 Chr6:113442118..113442252 CCCTCCGTGTTTTCACACTT Chr6:113442184..113442203 60.01 50
downstream ENSMUSE00000377202 Chr6:113442681..113443333 TTTCATCCTCCGTCATCTCC Chr6:113442722..113442741 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ACCTCTGAGCTACCCCTTCC Chr6:113437588..113437608 59.7 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACCTCTGAGCTACCCCTTCC Chr6:113437588..113437608 59.7 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000030284