Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI2122
Trapped Gene
Sfrs1 (ENSMUSG00000018379)
Vector Insertion
Chr 11: 87862544 - 87865111
Public Clones AE0827 (sanger) AE0837 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 74% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000674824 (Chr11:87862545..87867259 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000674824 (Chr11:87862545..87867259 +)
Downstram Exon
ENSMUSE00000379220 (Chr11:87862914..87865110 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000486255 Chr11:87861173..87861534 No primer for this exon
upstream ENSMUSE00000705979 Chr11:87861187..87861534 No primer for this exon
upstream ENSMUSE00000361135 Chr11:87861992..87862176 No primer for this exon

*** Putative Vector Insertion (Chr 11: 87862544 - 87865111) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000489619 Chr11:87862545..87862717 No primer for this exon
downstream ENSMUSE00000674824 Chr11:87862545..87867259 No primer for this exon
downstream ENSMUSE00000379220 Chr11:87862914..87865110 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTAGGGGGAATCGCTTTTTC Chr11:87862524..87862544 60.38 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTAGGGGGAATCGCTTTTTC Chr11:87862524..87862544 60.38 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000018379