Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI13261
Trapped Gene
Lasp1 (ENSMUSG00000038366)
Vector Insertion
Chr 11: 97677097 - 97686155
Public Clones (sanger) (sanger) CMHD-GT_340G9-3 (cmhd) PST17156-NR (escells) PST17178-NL (escells)
PST16924-NL (escells) PST17203-NL (escells) IST14862F7 (tigm) IST14806H3 (tigm)
IST14664H2 (tigm)
Private Clones OST358303 (lexicon) OST264358 (lexicon) OST51817 (lexicon) OST43592 (lexicon)
Severity of mutation (?) Insertion after 32% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000285046 (Chr11:97677012..97677096 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AACAGAGCGAGCTGCAGAGT Chr11:97677074..97677093 60.5 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000285046 (Chr11:97677012..97677096 +)
Downstram Exon
ENSMUSE00000673676 (Chr11:97686156..97686234 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AACAGAGCGAGCTGCAGAGT Chr11:97677074..97677093 60.5 55 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000673695 Chr11:97660983..97661176 CGGAACCATGAACCCTAACT Chr11:97661101..97661120 58.91 50
upstream ENSMUSE00000347026 Chr11:97660986..97661176 CGGAACCATGAACCCTAACT Chr11:97661101..97661120 58.91 50
upstream ENSMUSE00000673683 Chr11:97660989..97661176 CGGAACCATGAACCCTAACT Chr11:97661101..97661120 58.91 50
upstream ENSMUSE00000285055 Chr11:97668139..97668233 ATGCTTTCACTGCGAGACCT Chr11:97668153..97668172 60.02 50
upstream ENSMUSE00000673680 Chr11:97668139..97668243 ACTGCAATGCGTGAGTTTTG Chr11:97668224..97668243 59.91 45
upstream ENSMUSE00000673691 Chr11:97668139..97668233 ATGCTTTCACTGCGAGACCT Chr11:97668153..97668172 60.02 50
upstream ENSMUSE00000285046 Chr11:97677012..97677096 AACAGAGCGAGCTGCAGAGT Chr11:97677074..97677093 60.5 55
upstream ENSMUSE00000673679 Chr11:97677021..97677096 AACAGAGCGAGCTGCAGAGT Chr11:97677074..97677093 60.5 55

*** Putative Vector Insertion (Chr 11: 97677097 - 97686155) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000505619 Chr11:97686156..97686263 TCTGGTCCTGGGTCTTCTTG Chr11:97686258..97686277 60.23 55
downstream ENSMUSE00000673676 Chr11:97686156..97686234 No primer for this exon
downstream ENSMUSE00000673674 Chr11:97694738..97695016 ACTGGTCGGGATATGGTGAG Chr11:97695012..97695031 59.8 55
downstream ENSMUSE00000718694 Chr11:97694860..97695016 ACTGGTCGGGATATGGTGAG Chr11:97695012..97695031 59.8 55
downstream ENSMUSE00000722091 Chr11:97694860..97695016 ACTGGTCGGGATATGGTGAG Chr11:97695012..97695031 59.8 55
downstream ENSMUSE00000285023 Chr11:97695398..97695501 CCACCATAGGACGAGGTCAT Chr11:97695446..97695465 59.8 55
downstream ENSMUSE00000673673 Chr11:97695398..97695698 CCACCATAGGACGAGGTCAT Chr11:97695446..97695465 59.8 55
downstream ENSMUSE00000673689 Chr11:97695398..97695501 CCACCATAGGACGAGGTCAT Chr11:97695446..97695465 59.8 55
downstream ENSMUSE00000356471 Chr11:97697386..97700076 CCTCAGACACGACGACAGAA Chr11:97698772..97698791 60.02 55
downstream ENSMUSE00000673686 Chr11:97697386..97699291 CCTCAGACACGACGACAGAA Chr11:97698772..97698791 60.02 55
downstream ENSMUSE00000673684 Chr11:97700028..97700076 No primer for this exon
downstream ENSMUSE00000673675 Chr11:97736253..97736277 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ACCTGAACCCAAGCTCTCCT Chr11:97677119..97677139 60.25 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTTTGACGTGACTGGGAAAAC Chr11:97677142..97677163 60 42.86 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000038366