Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20493
Trapped Gene
Nus1 (ENSMUSG00000023068)
Vector Insertion
Chr 10: 52149125 - 52149872
Public Clones not available
Private Clones OST360652 (lexicon)
Severity of mutation (?) Insertion after 62% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000302205 (Chr10:52148999..52149124 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTTTTGGGCCAAGATTGTTC Chr10:52149058..52149077 59.55 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000302205 (Chr10:52148999..52149124 +)
Downstram Exon
ENSMUSE00000133320 (Chr10:52149873..52150022 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTTTTGGGCCAAGATTGTTC Chr10:52149058..52149077 59.55 45 GCTGCTGGGCTACTAACTGG Chr10:52149977..52149996 60.04 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000411272 Chr10:52137365..52137974 CTGTTGGTCACGGAGGAAGT Chr10:52137866..52137885 60.15 55
upstream ENSMUSE00000302205 Chr10:52148999..52149124 CTTTTGGGCCAAGATTGTTC Chr10:52149058..52149077 59.55 45

*** Putative Vector Insertion (Chr 10: 52149125 - 52149872) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000133320 Chr10:52149873..52150022 GCTGCTGGGCTACTAACTGG Chr10:52149977..52149996 60.04 60
downstream ENSMUSE00000133319 Chr10:52153151..52153250 CTGATTTGCCAGGGAAGAAA Chr10:52153238..52153257 60.18 45
downstream ENSMUSE00000362324 Chr10:52156383..52156570 GGCGGAGAAGAAGTCCTCAT Chr10:52156434..52156453 60.74 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCAAGGTAAGTGCGTGGACA Chr10:52149121..52149141 60.3 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCAAGGTAAGTGCGTGGACA Chr10:52149121..52149141 60.3 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023068