Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI2658
Trapped Gene
Ankle2 (ENSMUSG00000029501)
Vector Insertion
Chr 5: 110667155 - 110670947
Public Clones AL0586 (sanger) XP0058 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 47% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000298893 (Chr5:110666966..110667154 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGCGCATCCTCTACGTTGTT Chr5:110667105..110667124 59.9 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000298893 (Chr5:110666966..110667154 +)
Downstram Exon
ENSMUSE00000189570 (Chr5:110670948..110671070 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGCGCATCCTCTACGTTGTT Chr5:110667105..110667124 59.9 50 TCGGTTTTTCACAATCAAAGG Chr5:110671042..110671062 59.96 38.1

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000444655 Chr5:110660060..110660442 No primer for this exon
upstream ENSMUSE00000444642 Chr5:110663366..110663839 CTGTGGACGGGAAACAAACT Chr5:110663609..110663628 60.01 50
upstream ENSMUSE00000444634 Chr5:110665292..110665498 TGTCTCCTGTGAAGGCAGTG Chr5:110665451..110665470 60.02 55
upstream ENSMUSE00000540519 Chr5:110666674..110666870 TTACAAAAACCCCCGTACCC Chr5:110666729..110666748 60.8 50
upstream ENSMUSE00000298893 Chr5:110666966..110667154 AGCGCATCCTCTACGTTGTT Chr5:110667105..110667124 59.9 50

*** Putative Vector Insertion (Chr 5: 110667155 - 110670947) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000189570 Chr5:110670948..110671070 TCGGTTTTTCACAATCAAAGG Chr5:110671042..110671062 59.96 38.1
downstream ENSMUSE00000189574 Chr5:110673523..110673589 AGTGCTGGAGATTTGTTTTGG Chr5:110673562..110673582 59.23 42.86
downstream ENSMUSE00000189571 Chr5:110678350..110678519 TCTGGGGCTGCTTCTACAGT Chr5:110678471..110678490 60.01 55
downstream ENSMUSE00000189576 Chr5:110679720..110679826 TGCTTTCTTTCGAGGTGGAG Chr5:110679770..110679789 60.51 50
downstream ENSMUSE00000189577 Chr5:110680516..110680706 AGTCTTTGCAAGCCTTCCTG Chr5:110680614..110680633 59.62 50
downstream ENSMUSE00000298853 Chr5:110681731..110682361 TCAGTTGTGGGCTGACTCTG Chr5:110681860..110681879 60.02 55
downstream ENSMUSE00000540517 Chr5:110681731..110681816 TAGAAATGCTCCCACGGAGA Chr5:110681780..110681799 60.73 50
downstream ENSMUSE00000540514 Chr5:110682051..110682361 TTGAAGGAAGCACCTCATCC Chr5:110682153..110682172 60.19 50
downstream ENSMUSE00000189578 Chr5:110682802..110682933 TGTGGATTGCTGGGTACAGA Chr5:110682890..110682909 60.11 50
downstream ENSMUSE00000298834 Chr5:110683320..110685666 GATGGCTCGGGTAAAACTCA Chr5:110685445..110685464 60.07 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CGGATAAAGGCTTGAGTTCCT Chr5:110667179..110667200 59.73 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGACATTCGTGACTGGGAAA Chr5:110670199..110670219 60.09 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029501