Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI2526
Trapped Gene
Med27 (ENSMUSG00000026799)
Vector Insertion
Chr 2: 29205511 - 29268907
Public Clones XS0817 (sanger) XS0819 (sanger) RRS260 (baygenomics) IST12789F8 (tigm)
IST11115H11 (tigm) IST12947A11 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 56% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000162927 (Chr2:29205366..29205510 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATCCTGTGCAGGACAAAACC Chr2:29205446..29205465 59.97 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000162927 (Chr2:29205366..29205510 +)
Downstram Exon
ENSMUSE00000162923 (Chr2:29268908..29269038 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATCCTGTGCAGGACAAAACC Chr2:29205446..29205465 59.97 50 CAAAAGGCCAGACGCTAGAC Chr2:29268946..29268965 60.01 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000385559 Chr2:29202356..29202567 GCGTGAGTAGGGTCTTCGAC Chr2:29202444..29202463 59.87 60
upstream ENSMUSE00000162927 Chr2:29205366..29205510 ATCCTGTGCAGGACAAAACC Chr2:29205446..29205465 59.97 50

*** Putative Vector Insertion (Chr 2: 29205511 - 29268907) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000162923 Chr2:29268908..29269038 CAAAAGGCCAGACGCTAGAC Chr2:29268946..29268965 60.01 55
downstream ENSMUSE00000162915 Chr2:29326813..29326906 TCCCATTGGGTCTCGATAAG Chr2:29326892..29326911 59.89 50
downstream ENSMUSE00000697121 Chr2:29332853..29332906 AAGGGCCAGTAATGCTCTGA Chr2:29332895..29332914 59.84 50
downstream ENSMUSE00000162919 Chr2:29355739..29355846 GCTCCTCATGACCACAATGA Chr2:29355786..29355805 59.64 50
downstream ENSMUSE00000162917 Chr2:29364943..29364984 GGTAGCTGGATTTGGACCAT Chr2:29364973..29364992 58.86 50
downstream ENSMUSE00000162913 Chr2:29378294..29378371 ATGAAGGACCTGACCACGAC Chr2:29378373..29378392 59.97 55
downstream ENSMUSE00000360310 Chr2:29379887..29380308 TTCGGAAGTCTCTCCACGTC Chr2:29379989..29380008 60.39 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTTGGGTATCTTGCCCATGT Chr2:29247536..29247556 59.68 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTCCCTTCTTTGGTTCGTGA Chr2:29247547..29247567 60.22 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026799