Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI17948
Trapped Gene
Zfp462 (ENSMUSG00000060206)
Vector Insertion
Chr 4: 55036479 - 55063057
Public Clones IST13035C9 (tigm) IST11295D10 (tigm) IST10550D2 (tigm) IST10110F7 (tigm)
IST11411B4 (tigm) IST11084C12 (tigm) IST11382D4 (tigm)
Private Clones OST432890 (lexicon) OST44568 (lexicon)
Severity of mutation (?) Insertion after 74% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000179018 (Chr4:55036287..55036478 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAGCCGAAGAAGTGGAAGAC Chr4:55036452..55036471 59.99 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000179018 (Chr4:55036287..55036478 +)
Downstram Exon
ENSMUSE00000179012 (Chr4:55063058..55063325 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAGCCGAAGAAGTGGAAGAC Chr4:55036452..55036471 59.99 55 TAACGCGGCTCTTGTTTCTT Chr4:55063147..55063166 60.02 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000673985 Chr4:54960817..54961104 CAATAACCCGAGCAGGAAGA Chr4:54961008..54961027 60.21 50
upstream ENSMUSE00000487524 Chr4:55020405..55020654 TCCACACGGCATTCTTACAG Chr4:55020511..55020530 59.72 50
upstream ENSMUSE00000632627 Chr4:55021128..55026721 TTCATTCTGCGACAACTTGC Chr4:55022755..55022774 59.99 45
upstream ENSMUSE00000527230 Chr4:55024238..55026721 GACAAGCCCAACAAGGTGAT Chr4:55026086..55026105 59.97 50
upstream ENSMUSE00000632619 Chr4:55024352..55026721 GACAAGCCCAACAAGGTGAT Chr4:55026086..55026105 59.97 50
upstream ENSMUSE00000527229 Chr4:55027555..55027719 GGTGCCAAAGATCAAGGAGA Chr4:55027575..55027594 60.19 50
upstream ENSMUSE00000673956 Chr4:55029686..55029972 CTGCTTGCTTGATGCAATGT Chr4:55029781..55029800 60.02 45
upstream ENSMUSE00000673961 Chr4:55029689..55029972 CTGCTTGCTTGATGCAATGT Chr4:55029781..55029800 60.02 45
upstream ENSMUSE00000527228 Chr4:55029869..55029972 GCTGCCAGTATTGCTCCTTT Chr4:55029933..55029952 59.48 50
upstream ENSMUSE00000360129 Chr4:55032961..55033079 ACAGCCGGTTGAAGACACAC Chr4:55033041..55033060 61.18 55
upstream ENSMUSE00000179018 Chr4:55036287..55036478 CAGCCGAAGAAGTGGAAGAC Chr4:55036452..55036471 59.99 55

*** Putative Vector Insertion (Chr 4: 55036479 - 55063057) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000179012 Chr4:55063058..55063325 TAACGCGGCTCTTGTTTCTT Chr4:55063147..55063166 60.02 45
downstream ENSMUSE00000179016 Chr4:55064063..55064199 TGCAGCACTATGTGGTCGAT Chr4:55064197..55064216 60.3 50
downstream ENSMUSE00000179014 Chr4:55072886..55073109 AGTTTGGAACGGCACACTTC Chr4:55072928..55072947 60.16 50
downstream ENSMUSE00000179000 Chr4:55085845..55085977 GCTCATTTCGTCGTCCTTGT Chr4:55085961..55085980 60.26 50
downstream ENSMUSE00000179008 Chr4:55092136..55092259 CACGTGTCTTTCCCACTCAG Chr4:55092248..55092267 59.31 55
downstream ENSMUSE00000604453 Chr4:55093538..55096434 TAAACATGTGGCCAACGAAA Chr4:55094654..55094673 59.97 40
downstream ENSMUSE00000632620 Chr4:55093538..55094230 AGATCCGGAAGGAAGGATGT Chr4:55093992..55094011 59.9 50
downstream ENSMUSE00000673955 Chr4:55093538..55094202 AGATCCGGAAGGAAGGATGT Chr4:55093992..55094011 59.9 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTAGCGGCAGCTGTTTCTTT Chr4:55045494..55045514 59.66 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTAGCGGCAGCTGTTTCTTT Chr4:55045494..55045514 59.66 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000060206