Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI220
Trapped Gene
Myst4 (ENSMUSG00000021767)
Vector Insertion
Chr 14: 22456863 - 22479964
Public Clones GC0655 (tigem) CSD188 (baygenomics) HMA154 (baygenomics) CSJ235 (baygenomics)
Private Clones OST422981 (lexicon)
Severity of mutation (?) Insertion after 37% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000120701 (Chr14:22456769..22456862 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGGATTTGGACGCTTTCTCA Chr14:22456834..22456853 59.81 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000120701 (Chr14:22456769..22456862 +)
Downstram Exon
ENSMUSE00000120698 (Chr14:22479965..22480196 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGGATTTGGACGCTTTCTCA Chr14:22456834..22456853 59.81 45 TGGTGGCGGTAGAGGTATTC Chr14:22480091..22480110 59.96 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000377733 Chr14:22319076..22319182 No primer for this exon
upstream ENSMUSE00000650586 Chr14:22320107..22320300 ATGTCGGATTCATGTCAACG Chr14:22320117..22320136 59.37 45
upstream ENSMUSE00000352161 Chr14:22335848..22336720 TGGGTAACGTCTGTGTTGGA Chr14:22336060..22336079 60 50
upstream ENSMUSE00000691743 Chr14:22335848..22339526 TGGGTAACGTCTGTGTTGGA Chr14:22336060..22336079 60 50
upstream ENSMUSE00000691747 Chr14:22336097..22336507 TCTACGCAATGTGGATTGGA Chr14:22336402..22336421 60.07 45
upstream ENSMUSE00000120693 Chr14:22428976..22429084 CCGATCCCATTCCAATATGT Chr14:22428983..22429002 59.46 45
upstream ENSMUSE00000120710 Chr14:22438113..22438228 TGTCCTGAGTTAACCGCAAA Chr14:22438136..22438155 59.32 45
upstream ENSMUSE00000120702 Chr14:22438440..22438521 GTGACCCACCACTTTCCAGA Chr14:22438492..22438511 60.97 55
upstream ENSMUSE00000120708 Chr14:22441177..22441309 TAAAACGGCGATATGCAAAA Chr14:22441249..22441268 59.2 35
upstream ENSMUSE00000231429 Chr14:22444055..22444437 GCAGATCCCGAGGTGAAATA Chr14:22444339..22444358 60.04 50
upstream ENSMUSE00000231424 Chr14:22445941..22446062 GGAAACAAGGAGCTCGTGAC Chr14:22445994..22446013 59.85 55
upstream ENSMUSE00000231417 Chr14:22448035..22448150 CGGGAAATACGAGATCCAAA Chr14:22448094..22448113 59.89 45
upstream ENSMUSE00000120699 Chr14:22450544..22450685 CTCCGGCCAATGAGATTTAC Chr14:22450639..22450658 59.53 50
upstream ENSMUSE00000120703 Chr14:22453698..22453859 AAAGGCTGTCATCTGGTTGG Chr14:22453827..22453846 60.11 50
upstream ENSMUSE00000120701 Chr14:22456769..22456862 AGGATTTGGACGCTTTCTCA Chr14:22456834..22456853 59.81 45

*** Putative Vector Insertion (Chr 14: 22456863 - 22479964) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000120698 Chr14:22479965..22480196 TGGTGGCGGTAGAGGTATTC Chr14:22480091..22480110 59.96 55
downstream ENSMUSE00000231392 Chr14:22480650..22480809 TCTCAGCTTCTCGCTCTTCC Chr14:22480807..22480826 59.97 55
downstream ENSMUSE00000231383 Chr14:22481380..22481688 TTGCAGACGATTGCCTACTG Chr14:22481473..22481492 60.01 50
downstream ENSMUSE00000231374 Chr14:22483679..22483970 AGTTGTCGAAGGGCTCATTG Chr14:22483796..22483815 60.26 50
downstream ENSMUSE00000507785 Chr14:22487553..22491354 AGGGGCTTGTATCCACACTG Chr14:22490427..22490446 59.99 55
downstream ENSMUSE00000691746 Chr14:22489967..22490095 TGTGTTCATGTAGCCGTGGT Chr14:22490053..22490072 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GAGTCCCTGCCTCTTCACAG Chr14:22459852..22459872 59.99 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAGTCCCTGCCTCTTCACAG Chr14:22459852..22459872 59.99 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000021767