Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33161
Trapped Gene
Kcnc3 (ENSMUSG00000062785)
Vector Insertion
Chr 7: 51857238 - 51857512
Public Clones IST14932F6 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 100% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000513173 (Chr7:51857239..51857511 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAAAGAGACGATGCCAGCAG Chr7:51857488..51857507 60.54 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000513173 (Chr7:51857239..51857511 +)
Downstram Exon
ENSMUSE00000674786 (Chr7:51857239..51860124 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAAAGAGACGATGCCAGCAG Chr7:51857488..51857507 60.54 50 GAAGGGGCATGACACTGTTT Chr7:51860075..51860094 59.97 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000674785 Chr7:51846034..51847128 GCACGGACGAGTTCTTCTTC Chr7:51846653..51846672 60 55
upstream ENSMUSE00000491194 Chr7:51846256..51847128 GCACGGACGAGTTCTTCTTC Chr7:51846653..51846672 60 55
upstream ENSMUSE00000275186 Chr7:51850531..51851638 CCCTTCTTGACCTACGTGGA Chr7:51850696..51850715 60.1 55
upstream ENSMUSE00000275182 Chr7:51853751..51853945 GACAAGAGCCCAATCACTCC Chr7:51853840..51853859 59.66 55
upstream ENSMUSE00000674788 Chr7:51853754..51853945 GACAAGAGCCCAATCACTCC Chr7:51853840..51853859 59.66 55
upstream ENSMUSE00000674784 Chr7:51856239..51856298 CTTGTCACCGCCTGAGACCT Chr7:51856241..51856260 62.77 60

*** Putative Vector Insertion (Chr 7: 51857238 - 51857512) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000513173 Chr7:51857239..51857511 CCCTACTCCCAGCTCCTTTT Chr7:51857443..51857462 59.71 55
downstream ENSMUSE00000674786 Chr7:51857239..51860124 GAAGGGGCATGACACTGTTT Chr7:51860075..51860094 59.97 50
downstream ENSMUSE00000674787 Chr7:51857239..51857462 CCCTACTCCCAGCTCCTTTT Chr7:51857443..51857462 59.71 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCTGCTCCTTTCCCAGGTTA Chr7:51857224..51857244 59.81 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCTGCTCCTTTCCCAGGTTA Chr7:51857224..51857244 59.81 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000062785