Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI22504
Trapped Gene
Mrps18b (ENSMUSG00000024436)
Vector Insertion
Chr 17: 36051535 - 36051640
Public Clones PST22331-NR (escells)
Private Clones OST295042 (lexicon)
Severity of mutation (?) Insertion after 15% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000142322 (Chr17:36051641..36051749 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATATGAGAGCGAGCCTTGGA Chr17:36051659..36051678 59.94 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000142322 (Chr17:36051641..36051749 -)
Downstram Exon
ENSMUSE00000142318 (Chr17:36051437..36051534 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATATGAGAGCGAGCCTTGGA Chr17:36051659..36051678 59.94 50 GCCATAGCGGTTGTGGTATT Chr17:36051493..36051512 59.85 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000472037 Chr17:36053237..36053334 CGCTTCTAAAGCTGGTTCCT Chr17:36053267..36053286 58.74 50
upstream ENSMUSE00000142322 Chr17:36051641..36051749 ATATGAGAGCGAGCCTTGGA Chr17:36051659..36051678 59.94 50

*** Putative Vector Insertion (Chr 17: 36051535 - 36051640) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000142318 Chr17:36051437..36051534 GCCATAGCGGTTGTGGTATT Chr17:36051493..36051512 59.85 50
downstream ENSMUSE00000142320 Chr17:36051279..36051347 CAAGGATTCCCAGCAACTTT Chr17:36051297..36051316 59.17 45
downstream ENSMUSE00000142315 Chr17:36049283..36049349 GTGTATGGGGCGTGAAAGAT Chr17:36049263..36049282 59.82 50
downstream ENSMUSE00000142323 Chr17:36048364..36048423 CTGGGTCAGCTTCTTGTGCT Chr17:36048367..36048386 60.59 55
downstream ENSMUSE00000475086 Chr17:36047331..36047901 CCTTGATAGAGTCGGCGAAG Chr17:36047696..36047715 59.97 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTGAGGCTAGGGGAAAGGAG Chr17:36051615..36051635 60.33 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGAGGCTAGGGGAAAGGAG Chr17:36051615..36051635 60.33 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GGTTCCCCTCCAGACTCTTT Chr17:36051729..36051749 59.53 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 GGTTCCCCTCCAGACTCTTT Chr17:36051729..36051749 59.53 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024436