Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI2668
Trapped Gene
Mrpl20 (ENSMUSG00000029066)
Vector Insertion
Chr 4: 155177852 - 155177941
Public Clones AL0553 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 19% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000185188 (Chr4:155177743..155177851 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CATCATGGTCTTCCTCACGA Chr4:155177761..155177780 59.63 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000185188 (Chr4:155177743..155177851 +)
Downstram Exon
ENSMUSE00000185186 (Chr4:155177942..155178052 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CATCATGGTCTTCCTCACGA Chr4:155177761..155177780 59.63 50 CGGGCCTTCGTACACTTAAC Chr4:155178027..155178046 59.63 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000185188 Chr4:155177743..155177851 CATCATGGTCTTCCTCACGA Chr4:155177761..155177780 59.63 50

*** Putative Vector Insertion (Chr 4: 155177852 - 155177941) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000185186 Chr4:155177942..155178052 CGGGCCTTCGTACACTTAAC Chr4:155178027..155178046 59.63 55
downstream ENSMUSE00000185187 Chr4:155181017..155181094 GGAGGCAGCTGTAATTCGAT Chr4:155181049..155181068 59.3 50
downstream ENSMUSE00000228704 Chr4:155182588..155182938 TGGTACTGCACCACTCTGGA Chr4:155182760..155182779 60.31 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTGCTGAAACACGCTCAGGT Chr4:155177835..155177855 61.46 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGCTGAAACACGCTCAGGT Chr4:155177835..155177855 61.46 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029066