Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32898
Trapped Gene
Eepd1 (ENSMUSG00000036611)
Vector Insertion
Chr 9: 25355189 - 25389476
Public Clones IST11996A3 (tigm) IST10588G11 (tigm) IST12443F4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 54% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000268158 (Chr9:25355137..25355188 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACAAAGAGGCCCTGGAAAAG Chr9:25355169..25355188 60.6 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000268158 (Chr9:25355137..25355188 +)
Downstram Exon
ENSMUSE00000268127 (Chr9:25389477..25389587 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACAAAGAGGCCCTGGAAAAG Chr9:25355169..25355188 60.6 50 AACAATGGACCTCCAGCATC Chr9:25389560..25389579 59.93 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000413905 Chr9:25289182..25289481 CTGGATTCAAGCAGCTACGA Chr9:25289287..25289306 59.17 50
upstream ENSMUSE00000410963 Chr9:25289866..25290904 GAATATATCGGCGGCTTCAA Chr9:25290231..25290250 60.03 45
upstream ENSMUSE00000268158 Chr9:25355137..25355188 ACAAAGAGGCCCTGGAAAAG Chr9:25355169..25355188 60.6 50

*** Putative Vector Insertion (Chr 9: 25355189 - 25389476) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000268127 Chr9:25389477..25389587 AACAATGGACCTCCAGCATC Chr9:25389560..25389579 59.93 50
downstream ENSMUSE00000268098 Chr9:25394247..25394381 CTTGAATCTGGCGAGGAAAG Chr9:25394384..25394403 59.95 50
downstream ENSMUSE00000268075 Chr9:25397020..25397158 TCCTGGAGGGTTAGTGCAAA Chr9:25397150..25397169 60.63 50
downstream ENSMUSE00000268052 Chr9:25402146..25402340 GAAAGTGTGAGCCGGGACTA Chr9:25402258..25402277 60.26 55
downstream ENSMUSE00000331342 Chr9:25410994..25411694 CCCTGTAGTGCCATCTCCAT Chr9:25411257..25411276 59.95 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTGTGACCTGATGGGGACTT Chr9:25376166..25376186 59.96 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAAGACCGTGACTGGGAAAA Chr9:25376234..25376254 61.06 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000036611