Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI37640
Trapped Gene
Sertad2 (ENSMUSG00000049800)
Vector Insertion
Chr 11: 20547804 - 20549069
Public Clones (cmhd)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000680840 (Chr11:20547805..20549068 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACACCTTACAGCGCCAGACT Chr11:20547906..20547925 59.94 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000680840 (Chr11:20547805..20549068 +)
Downstram Exon
ENSMUSE00000580920 (Chr11:20547805..20553026 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACACCTTACAGCGCCAGACT Chr11:20547906..20547925 59.94 55 AAGCAACCAGATTGGACACC Chr11:20550326..20550345 59.97 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000593852 Chr11:20443256..20443763 GTATCGCCCCTTTAGCCTGT Chr11:20443494..20443513 60.47 55
upstream ENSMUSE00000680843 Chr11:20531980..20532276 CTGGAGCACGAGCTGTACCT Chr11:20532019..20532038 60.61 60
upstream ENSMUSE00000680841 Chr11:20547158..20547355 GATACCCGACAGGCAAGAAG Chr11:20547276..20547295 59.69 55

*** Putative Vector Insertion (Chr 11: 20547804 - 20549069) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000580920 Chr11:20547805..20553026 AAGCAACCAGATTGGACACC Chr11:20550326..20550345 59.97 50
downstream ENSMUSE00000680840 Chr11:20547805..20549068 GGGGTCAAAATCGTACATGG Chr11:20548576..20548595 60.05 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGCCTTCATGTGATTCATGC Chr11:20547761..20547781 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGATCGTGACTGGGAAAACC Chr11:20547851..20547871 58.99 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000049800