Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14564
Trapped Gene
Hsp90ab1 (ENSMUSG00000023944)
Vector Insertion
Chr 17: 45704727 - 45705512
Public Clones CMHD-GT_396B9-3 (cmhd) FHCRC-GT-S6-11C1 (fhcrc)
Private Clones OST181473 (lexicon) OST171790 (lexicon) OST107360 (lexicon)
Severity of mutation (?) Insertion after 95% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000136561 (Chr17:45705178..45705511 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACAAGCACCTATGGCTGGAC Chr17:45705447..45705466 60.14 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000136561 (Chr17:45705178..45705511 -)
Downstram Exon
ENSMUSE00000657294 (Chr17:45704728..45705073 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACAAGCACCTATGGCTGGAC Chr17:45705447..45705466 60.14 55 TTCCAGGAGGCTTTAATCCA Chr17:45704930..45704949 59.64 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000696089 Chr17:45710116..45710220 CCACCACCCTGCTCTGTACT Chr17:45710138..45710157 60.17 60
upstream ENSMUSE00000136555 Chr17:45708714..45708860 AAGTGCACCATGGAGAGGAG Chr17:45708831..45708850 60.26 55
upstream ENSMUSE00000404358 Chr17:45708362..45708568 GCCCTGGACAAGATTCGATA Chr17:45708549..45708568 60.04 50
upstream ENSMUSE00000136547 Chr17:45707967..45708126 GCAGTTTGGTGTCGGATTCT Chr17:45708081..45708100 60.12 50
upstream ENSMUSE00000136551 Chr17:45707615..45707748 AGGAGGGTCAAGGAAGTGGT Chr17:45707658..45707677 59.97 55
upstream ENSMUSE00000136553 Chr17:45707207..45707515 ATGACTGGGAGGACCACTTG Chr17:45707216..45707235 59.96 55
upstream ENSMUSE00000136557 Chr17:45706419..45706775 AGGTCATCCGCAAGAACATC Chr17:45706509..45706528 60.08 50
upstream ENSMUSE00000136550 Chr17:45706189..45706336 TGAGCTCCTTCGCTATCACA Chr17:45706276..45706295 59.7 50
upstream ENSMUSE00000543014 Chr17:45705807..45706075 GAGCTACCAGAGGACGAGGA Chr17:45705895..45705914 59.56 60
upstream ENSMUSE00000136561 Chr17:45705178..45705511 ACAAGCACCTATGGCTGGAC Chr17:45705447..45705466 60.14 55
upstream ENSMUSE00000657294 Chr17:45704728..45705073 AAGATCCCCTCCCACAGAAT Chr17:45704819..45704838 59.76 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCACAGGTGACAATCTCCAA Chr17:45705496..45705516 59.52 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGACAACGTGACTGGGAAAA Chr17:45705447..45705467 60.13 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GGGTGAGTCTGGCTTGTGAG Chr17:45705158..45705178 60.86 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 ACTAGCTGGACCGTGACTGG Chr17:45705117..45705137 60.32 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023944