Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31040
Trapped Gene
Fhit (ENSMUSG00000060579)
Vector Insertion
Chr 14: 10702543 - 11254112
Public Clones (sanger) IST14154C12 (tigm) IST10528H3 (tigm) IST14257B11 (tigm) IST13735E11 (tigm)
IST14857D1 (tigm) IST14773F8 (tigm) IST10320H9 (tigm) IST14542H3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000565183 (Chr14:11254009..11254111 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTCATCAAGCCCTCTGTGGT Chr14:11254068..11254087 60.26 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000565183 (Chr14:11254009..11254111 -)
Downstram Exon
ENSMUSE00000444519 (Chr14:10702544..10702689 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTCATCAAGCCCTCTGTGGT Chr14:11254068..11254087 60.26 55 AACAAATCGGCCACTTCATC Chr14:10702595..10702614 59.94 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000565183 Chr14:11254009..11254111 CTCATCAAGCCCTCTGTGGT Chr14:11254068..11254087 60.26 55
upstream ENSMUSE00000444519 Chr14:10702544..10702689 GTGGCCGATTTGTTTCAAGT Chr14:10702611..10702630 59.98 45
upstream ENSMUSE00000692685 Chr14:10702544..10702689 GTGGCCGATTTGTTTCAAGT Chr14:10702611..10702630 59.98 45

*** Putative Vector Insertion (Chr 14: 10702543 - 11254112) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000444509 Chr14:10694531..10694560 CTTCACAGTCTGCCCAGCTT Chr14:10694509..10694528 60.59 55
downstream ENSMUSE00000692683 Chr14:10694446..10694451 No primer for this exon
downstream ENSMUSE00000565180 Chr14:10596254..10596322 No primer for this exon
downstream ENSMUSE00000565178 Chr14:10405924..10406025 CCTGAAAGTAGACCCGCAGA Chr14:10405904..10405923 60.39 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCACAGCTCTTGCCCTGAAT Chr14:10753096..10753116 60.94 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCACAGCTCTTGCCCTGAAT Chr14:10753096..10753116 60.94 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000060579