Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI4697
Trapped Gene
Numa1 (ENSMUSG00000066306)
Vector Insertion
Chr 7: 109144450 - 109144536
Public Clones AC0064 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 14% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000633241 (Chr7:109144332..109144449 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTCCGAGGCAAGAATGAAAG Chr7:109144430..109144449 59.95 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000633241 (Chr7:109144332..109144449 +)
Downstram Exon
ENSMUSE00000633240 (Chr7:109144537..109144654 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTCCGAGGCAAGAATGAAAG Chr7:109144430..109144449 59.95 50 GATCCATCTGGCTCTTCTCG Chr7:109144610..109144629 59.91 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000671692 Chr7:109118357..109118483 GCAGCTATCGGGACAGAAGA Chr7:109118358..109118377 60.5 55
upstream ENSMUSE00000528838 Chr7:109125894..109125964 CAGCTACGCTCCTTTCTTGG Chr7:109125945..109125964 60.15 55
upstream ENSMUSE00000633252 Chr7:109136240..109136325 CAGGACTGCAGCATCTTCAT Chr7:109136288..109136307 58.99 50
upstream ENSMUSE00000633251 Chr7:109139087..109139166 GAGGGGCAACAAATCCTACA Chr7:109139100..109139119 59.93 50
upstream ENSMUSE00000633250 Chr7:109140160..109140242 TAGAGGGATCCGAGATGGAA Chr7:109140214..109140233 59.58 50
upstream ENSMUSE00000633247 Chr7:109141152..109141232 CCATGAGCTCCAGAAACCTC Chr7:109141180..109141199 59.8 55
upstream ENSMUSE00000633245 Chr7:109141739..109141826 GCTGAGTTAGCGGTCATCCT Chr7:109141739..109141758 59.46 55
upstream ENSMUSE00000633244 Chr7:109143177..109143294 CCAAGAGGAAGATTCGCTTC Chr7:109143234..109143253 59.01 50
upstream ENSMUSE00000633242 Chr7:109143911..109144068 CAGACCCCACAGTTCCAGAT Chr7:109143954..109143973 59.96 55
upstream ENSMUSE00000633241 Chr7:109144332..109144449 CTCCGAGGCAAGAATGAAAG Chr7:109144430..109144449 59.95 50

*** Putative Vector Insertion (Chr 7: 109144450 - 109144536) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000633240 Chr7:109144537..109144654 GATCCATCTGGCTCTTCTCG Chr7:109144610..109144629 59.91 55
downstream ENSMUSE00000633239 Chr7:109144949..109145089 TTGAATGCACCCTGAAGTTG Chr7:109144998..109145017 59.69 45
downstream ENSMUSE00000633238 Chr7:109146589..109146711 CAGCCGAGTTGCTTGATCTT Chr7:109146663..109146682 60.54 50
downstream ENSMUSE00000528763 Chr7:109146814..109150173 TGTGCTTCTTGTCGCATTTC Chr7:109150064..109150083 59.99 45
downstream ENSMUSE00000528759 Chr7:109156000..109156068 TGGTCATGCTCTGTCAGCTT Chr7:109156040..109156059 59.58 50
downstream ENSMUSE00000528756 Chr7:109157605..109157724 GACCTCTTGCTGGCTTTCAC Chr7:109157646..109157665 60 55
downstream ENSMUSE00000528753 Chr7:109157808..109158026 GTTCCTGCAGCTTTTGGTTC Chr7:109157871..109157890 59.86 50
downstream ENSMUSE00000528751 Chr7:109158131..109158288 TCACGGCTCTTTAAGGCATC Chr7:109158207..109158226 60.35 50
downstream ENSMUSE00000528748 Chr7:109159347..109159593 ACCATCTGGTTGGGTACGAG Chr7:109159377..109159396 59.84 55
downstream ENSMUSE00000528744 Chr7:109160395..109160623 CTGCGTGCTGTAGAAGGATG Chr7:109160451..109160470 59.62 55
downstream ENSMUSE00000528740 Chr7:109161256..109161392 TGGACTCCAGAGGGTAGCAG Chr7:109161393..109161412 60.4 60
downstream ENSMUSE00000528736 Chr7:109161683..109161859 CTGGCTCTACGCAAGGTTTC Chr7:109161762..109161781 60.01 55
downstream ENSMUSE00000528733 Chr7:109162044..109162160 GCTGCTCTGCTTACGTCCTT Chr7:109162121..109162140 59.79 55
downstream ENSMUSE00000528729 Chr7:109162252..109162455 TTGAAGCTCCTCGCCTTAGA Chr7:109162339..109162358 60.23 50
downstream ENSMUSE00000671673 Chr7:109162750..109163472 ACCCCTGAGATCCAGAACCT Chr7:109163321..109163340 59.93 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TATGACTCTCCCTGCCATCC Chr7:109144452..109144472 60.03 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGCTAGCGAGGTCGTGACT Chr7:109144488..109144508 60.75 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000066306