Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI15422
Trapped Gene
Lyrm5 (ENSMUSG00000040370)
Vector Insertion
Chr 6: 145159997 - 145163041
Public Clones (sanger) (sanger) (ggtc) PST4848-NR (escells) PST7097-NR (escells)
IST14914A2 (tigm) IST14913A9 (tigm) IST14914A2 (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
ENSMUSE00000688767 (Chr6:145159654..145159996 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGAGCTGCGTCTCACACTGA Chr6:145159806..145159825 59.92 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000688767 (Chr6:145159654..145159996 +)
Downstram Exon
ENSMUSE00000688752 (Chr6:145163042..145163065 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGAGCTGCGTCTCACACTGA Chr6:145159806..145159825 59.92 55 ATGCTTTTGACTCGGTGCTT Chr6:145163067..145163086 59.88 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000688767 Chr6:145159654..145159996 AGAGCTGCGTCTCACACTGA Chr6:145159806..145159825 59.92 55
upstream ENSMUSE00000505978 Chr6:145159676..145159772 TCTGAGCAAGCTGTCGTCTG Chr6:145159729..145159748 60.49 55
upstream ENSMUSE00000688759 Chr6:145159695..145159764 TCTGAGCAAGCTGTCGTCTG Chr6:145159729..145159748 60.49 55
upstream ENSMUSE00000688765 Chr6:145159695..145159772 TCTGAGCAAGCTGTCGTCTG Chr6:145159729..145159748 60.49 55
upstream ENSMUSE00000688757 Chr6:145159706..145159764 TCTGAGCAAGCTGTCGTCTG Chr6:145159729..145159748 60.49 55
upstream ENSMUSE00000688754 Chr6:145159720..145159772 TCTGAGCAAGCTGTCGTCTG Chr6:145159729..145159748 60.49 55

*** Putative Vector Insertion (Chr 6: 145159997 - 145163041) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000280866 Chr6:145163042..145163791 ATGCTTTTGACTCGGTGCTT Chr6:145163067..145163086 59.88 45
downstream ENSMUSE00000688752 Chr6:145163042..145163065 ATGCTTTTGACTCGGTGCTT Chr6:145163067..145163086 59.88 45
downstream ENSMUSE00000688763 Chr6:145163042..145163304 ATGCTTTTGACTCGGTGCTT Chr6:145163067..145163086 59.88 45
downstream ENSMUSE00000688751 Chr6:145163285..145163791 AACGAATTGGCCATTTTCAC Chr6:145163763..145163782 59.81 40
downstream ENSMUSE00000688769 Chr6:145163704..145163791 AACGAATTGGCCATTTTCAC Chr6:145163763..145163782 59.81 40
downstream ENSMUSE00000710310 Chr6:145163704..145163791 AACGAATTGGCCATTTTCAC Chr6:145163763..145163782 59.81 40
downstream ENSMUSE00000280858 Chr6:145163871..145165459 ACTGGTGCAATGATTGGTCA Chr6:145164106..145164125 59.97 45
downstream ENSMUSE00000688750 Chr6:145163871..145164254 TTGGATAGTCCCGTCCAAGA Chr6:145163901..145163920 60.45 50
downstream ENSMUSE00000688758 Chr6:145163871..145164291 TTGGATAGTCCCGTCCAAGA Chr6:145163901..145163920 60.45 50
downstream ENSMUSE00000688762 Chr6:145163871..145165059 ACTGGTGCAATGATTGGTCA Chr6:145164106..145164125 59.97 45
downstream ENSMUSE00000688766 Chr6:145163871..145164311 TTGGATAGTCCCGTCCAAGA Chr6:145163901..145163920 60.45 50
downstream ENSMUSE00000688768 Chr6:145163871..145165056 ACTGGTGCAATGATTGGTCA Chr6:145164106..145164125 59.97 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAGCCATACTGGATGGAACA Chr6:145162968..145162988 59.52 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAGCCATACTGGATGGAACA Chr6:145162968..145162988 59.52 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000040370