Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25326
Trapped Gene
Tnk2 (ENSMUSG00000022791)
Vector Insertion
Chr 16: 32673367 - 32675607
Public Clones CMHD-GT_421D7-3 (cmhd) (cmhd)
Private Clones OST187772 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000701757 (Chr16:32673242..32673366 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGGTGCCAGCTTTTACTCG Chr16:32673319..32673338 60.01 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000701757 (Chr16:32673242..32673366 +)
Downstram Exon
ENSMUSE00000701756 (Chr16:32675608..32675802 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGGTGCCAGCTTTTACTCG Chr16:32673319..32673338 60.01 55 ATTTCGAGGGAAGGGTCCTA Chr16:32675677..32675696 59.9 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000701777 Chr16:32644729..32645194 AGTCTGAGCATCCTCGAAGC Chr16:32645144..32645163 59.71 55
upstream ENSMUSE00000701762 Chr16:32644759..32645194 AGTCTGAGCATCCTCGAAGC Chr16:32645144..32645163 59.71 55
upstream ENSMUSE00000701761 Chr16:32644912..32645194 CATCCTCGAAGCAGTGTCTG Chr16:32645152..32645171 59.57 55
upstream ENSMUSE00000701759 Chr16:32644947..32645194 CATCCTCGAAGCAGTGTCTG Chr16:32645152..32645171 59.57 55
upstream ENSMUSE00000619840 Chr16:32663825..32664005 ACCTCAACATTACCCGCCTA Chr16:32663928..32663947 59.45 50
upstream ENSMUSE00000717799 Chr16:32663825..32664005 ACCTCAACATTACCCGCCTA Chr16:32663928..32663947 59.45 50
upstream ENSMUSE00000619839 Chr16:32665972..32666042 CATGTGCAAACGCAAGTCAT Chr16:32666012..32666031 60.73 45
upstream ENSMUSE00000619838 Chr16:32668307..32668528 GCACTCTCAGAGCACCTTCC Chr16:32668348..32668367 60.14 60
upstream ENSMUSE00000462682 Chr16:32669473..32669625 CATTCGCTTGTATGGTGTGG Chr16:32669583..32669602 59.99 50
upstream ENSMUSE00000130455 Chr16:32670039..32670316 AAGATTGGGGACTTCGGACT Chr16:32670228..32670247 59.94 50
upstream ENSMUSE00000130459 Chr16:32670878..32671004 GCCTGAAGACACGGACTTTC Chr16:32670892..32670911 59.85 55
upstream ENSMUSE00000130465 Chr16:32671381..32671527 CAATGTCATGGTCCAGTGCT Chr16:32671449..32671468 59.55 50
upstream ENSMUSE00000130457 Chr16:32671625..32671719 GCACATCCAGATGAATGACG Chr16:32671678..32671697 60.08 50
upstream ENSMUSE00000701757 Chr16:32673242..32673366 GAGGTGCCAGCTTTTACTCG Chr16:32673319..32673338 60.01 55

*** Putative Vector Insertion (Chr 16: 32673367 - 32675607) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000130468 Chr16:32675608..32675802 ATTTCGAGGGAAGGGTCCTA Chr16:32675677..32675696 59.9 50
downstream ENSMUSE00000701756 Chr16:32675608..32675802 ATTTCGAGGGAAGGGTCCTA Chr16:32675677..32675696 59.9 50
downstream ENSMUSE00000130470 Chr16:32677980..32678071 TTCCACACTCAGCAGGTCAG Chr16:32678031..32678050 60.02 55
downstream ENSMUSE00000701755 Chr16:32677980..32678071 TTCCACACTCAGCAGGTCAG Chr16:32678031..32678050 60.02 55
downstream ENSMUSE00000130454 Chr16:32678755..32678799 GAGTGAAGATGGCAGGCTGA Chr16:32678799..32678818 61.52 55
downstream ENSMUSE00000130471 Chr16:32679545..32680893 TAGAGAGCCGATGCGGTAGT Chr16:32680429..32680448 60 55
downstream ENSMUSE00000130473 Chr16:32680986..32681081 CAGCCCTCTGAACACTCCAG Chr16:32681070..32681089 61 60
downstream ENSMUSE00000515017 Chr16:32681238..32681365 CTAGGACCTTGTGGCACTCC Chr16:32681298..32681317 59.72 60
downstream ENSMUSE00000701758 Chr16:32681536..32681539 No primer for this exon
downstream ENSMUSE00000701760 Chr16:32682771..32683578 CACTGCTCTCCAGTCAACCA Chr16:32682800..32682819 60.02 55
downstream ENSMUSE00000701763 Chr16:32682771..32683579 CACTGCTCTCCAGTCAACCA Chr16:32682800..32682819 60.02 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGGAGGTGAGAGACATGAGC Chr16:32673363..32673383 59.8 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGGAGGTGAGAGACATGAGC Chr16:32673363..32673383 59.8 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022791