Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20759
Trapped Gene
Siglec5 (ENSMUSG00000039013)
Vector Insertion
Chr 7: 50607380 - 50607537
Public Clones not available
Private Clones OST352929 (lexicon)
Severity of mutation (?) Insertion after 25% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000281718 (Chr7:50607017..50607379 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGACGGTTATTCGCTCTCAG Chr7:50607030..50607049 59.84 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000281718 (Chr7:50607017..50607379 +)
Downstram Exon
ENSMUSE00000199270 (Chr7:50607538..50607816 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGACGGTTATTCGCTCTCAG Chr7:50607030..50607049 59.84 55 AATTGCCAGACACCAGGGTA Chr7:50607594..50607613 60.38 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000635168 Chr7:50606765..50606790 No primer for this exon
upstream ENSMUSE00000712027 Chr7:50606778..50606954 TGACATTGAAGCCTGTGACC Chr7:50606817..50606836 59.68 50
upstream ENSMUSE00000384161 Chr7:50606894..50606954 No primer for this exon
upstream ENSMUSE00000714426 Chr7:50606918..50606954 No primer for this exon
upstream ENSMUSE00000281718 Chr7:50607017..50607379 GGACGGTTATTCGCTCTCAG Chr7:50607030..50607049 59.84 55

*** Putative Vector Insertion (Chr 7: 50607380 - 50607537) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000199270 Chr7:50607538..50607816 AATTGCCAGACACCAGGGTA Chr7:50607594..50607613 60.38 50
downstream ENSMUSE00000281704 Chr7:50607992..50608039 TGGTCATCTTCTGTGGAGCA Chr7:50608015..50608034 60.4 50
downstream ENSMUSE00000281695 Chr7:50610346..50610597 AGGGACTCACCCTCTTGGAT Chr7:50610403..50610422 59.93 55
downstream ENSMUSE00000281688 Chr7:50610697..50610999 GGAGCTGGACTTAACCGTGA Chr7:50610863..50610882 60.26 55
downstream ENSMUSE00000199264 Chr7:50611270..50611369 CAAGGCCATAAGACCAGCTC Chr7:50611340..50611359 59.84 55
downstream ENSMUSE00000199248 Chr7:50611671..50611806 CAACGCAATGTTGGATGAAG Chr7:50611794..50611813 60.11 45
downstream ENSMUSE00000711625 Chr7:50611935..50611988 ACGTTGGGAAGAGCTTTGCT Chr7:50611989..50612008 61.31 50
downstream ENSMUSE00000712635 Chr7:50611935..50611988 ACGTTGGGAAGAGCTTTGCT Chr7:50611989..50612008 61.31 50
downstream ENSMUSE00000376892 Chr7:50613953..50614838 TTTGGTTTAGAGGGCCACTG Chr7:50614255..50614274 60.1 50
downstream ENSMUSE00000711723 Chr7:50613953..50614901 TTTGGTTTAGAGGGCCACTG Chr7:50614255..50614274 60.1 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGGATATGTGGAGGTCCTTG Chr7:50607393..50607413 59.21 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGGATATGTGGAGGTCCTTG Chr7:50607393..50607413 59.21 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000039013