Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI5712
Trapped Gene
Zfp462 (ENSMUSG00000060206)
Vector Insertion
Chr 4: 55064200 - 55072885
Public Clones CSI042 (baygenomics) IST11419H1 (tigm) IST11419H1 (tigm) IST10815A12 (tigm)
Private Clones OST364177 (lexicon) OST32050 (lexicon)
Severity of mutation (?) Insertion after 84% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000179016 (Chr4:55064063..55064199 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACAAACGCAACATGATCGAC Chr4:55064161..55064180 59.58 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000179016 (Chr4:55064063..55064199 +)
Downstram Exon
ENSMUSE00000179014 (Chr4:55072886..55073109 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACAAACGCAACATGATCGAC Chr4:55064161..55064180 59.58 45 AGTTTGGAACGGCACACTTC Chr4:55072928..55072947 60.16 50

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
upstream ENSMUSE00000179012 Chr4:55063058..55063325 AAGAAACAAGAGCCGCGTTA Chr4:55063125..55063144 60.02 45
upstream ENSMUSE00000179016 Chr4:55064063..55064199 ACAAACGCAACATGATCGAC Chr4:55064161..55064180 59.58 45

*** Putative Vector Insertion (Chr 4: 55064200 - 55072885) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
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 CCAGAGGGCCCTAAGTTTCT Chr4:55064170..55064190 59.71 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGAAAGCCTGGTCTTCGTGA Chr4:55064236..55064256 59.99 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000060206