Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI3479
Trapped Gene
Myo10 (ENSMUSG00000022272)
Vector Insertion
Chr 15: 25674012 - 25682742
Public Clones XQ0028 (sanger) AX0462 (sanger) H005H04 (ggtc) H004D02 (ggtc) P104A12 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000562992 (Chr15:25673931..25674011 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGGCAACACTAAGCTCCTCA Chr15:25673950..25673969 59.59 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000562992 (Chr15:25673931..25674011 +)
Downstram Exon
ENSMUSE00000684000 (Chr15:25682743..25683075 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGGCAACACTAAGCTCCTCA Chr15:25673950..25673969 59.59 50 TTGACCATAGGTGCCATTGA Chr15:25682835..25682854 59.92 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000624397 Chr15:25552280..25552785 GTTTGCTGGTGATCCGAAGT Chr15:25552371..25552390 60.12 50
upstream ENSMUSE00000684011 Chr15:25595399..25595411 No primer for this exon
upstream ENSMUSE00000684010 Chr15:25596176..25596279 CACGGGTCTGGCTAAGAGAA Chr15:25596185..25596204 60.39 55
upstream ENSMUSE00000451753 Chr15:25596181..25596279 CACGGGTCTGGCTAAGAGAA Chr15:25596185..25596204 60.39 55
upstream ENSMUSE00000563015 Chr15:25631317..25631475 AGCGCTACAAAAGGAACCAA Chr15:25631450..25631469 59.88 45
upstream ENSMUSE00000563013 Chr15:25643784..25643971 AACGAATGCTATCGCTGCTT Chr15:25643916..25643935 60.01 45
upstream ENSMUSE00000563011 Chr15:25651925..25652059 AGTTCCTGTCCGTCATCAGC Chr15:25651972..25651991 60.27 55
upstream ENSMUSE00000464787 Chr15:25653639..25653763 TTGGCAATGCGAAGACAGTA Chr15:25653656..25653675 60.4 45
upstream ENSMUSE00000649969 Chr15:25654827..25654840 No primer for this exon
upstream ENSMUSE00000486227 Chr15:25654951..25655035 CCGGGGAGAGGAATTATCAC Chr15:25654973..25654992 60.65 55
upstream ENSMUSE00000483446 Chr15:25656175..25656278 TGGGTGTACGGAAGACAAGA Chr15:25656224..25656243 59.13 50
upstream ENSMUSE00000484558 Chr15:25661725..25661854 TGCATCTCGGCAACATAGAG Chr15:25661795..25661814 59.97 50
upstream ENSMUSE00000489860 Chr15:25663743..25663861 GAGATCCTGACGCCTCTCAG Chr15:25663832..25663851 60.1 60
upstream ENSMUSE00000563006 Chr15:25665701..25665847 ACTTCAAATCCATCGGCATC Chr15:25665798..25665817 59.9 45
upstream ENSMUSE00000563004 Chr15:25666156..25666256 TGCAAATGAAAAGCTTCAGG Chr15:25666188..25666207 59.04 40
upstream ENSMUSE00000563002 Chr15:25666350..25666416 GGGATTGGTGTGGGAAGATA Chr15:25666353..25666372 59.6 50
upstream ENSMUSE00000563001 Chr15:25667205..25667297 CCATTTTCCTCAAGCCACAG Chr15:25667240..25667259 60.63 50
upstream ENSMUSE00000562998 Chr15:25667712..25667780 TCAACAATTTTGGCGTGAAG Chr15:25667746..25667765 59.71 40
upstream ENSMUSE00000562997 Chr15:25669042..25669124 CGAGGCATCTTGGAAAAGAA Chr15:25669057..25669076 60.32 45
upstream ENSMUSE00000562995 Chr15:25672096..25672204 CTGTCAGCTCGCAGTTCAAG Chr15:25672185..25672204 59.92 55
upstream ENSMUSE00000562992 Chr15:25673931..25674011 TGGCAACACTAAGCTCCTCA Chr15:25673950..25673969 59.59 50

*** Putative Vector Insertion (Chr 15: 25674012 - 25682742) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000624376 Chr15:25682665..25683075 TTGACCATAGGTGCCATTGA Chr15:25682835..25682854 59.92 45
downstream ENSMUSE00000684000 Chr15:25682743..25683075 TTGACCATAGGTGCCATTGA Chr15:25682835..25682854 59.92 45
downstream ENSMUSE00000562961 Chr15:25705776..25705900 ATACCCTGCTTTCCGAATCC Chr15:25705868..25705887 60.29 50
downstream ENSMUSE00000562960 Chr15:25706073..25706187 TCATAGACCTGCAGGAGCAC Chr15:25706153..25706172 58.99 55
downstream ENSMUSE00000649963 Chr15:25707836..25707942 GCACGGTCAATCTCCTCTTC Chr15:25707903..25707922 59.81 55
downstream ENSMUSE00000649961 Chr15:25709338..25709565 ACCACACCACAGAGGACCTT Chr15:25709373..25709392 59.45 55
downstream ENSMUSE00000649959 Chr15:25710119..25710167 GGAGGTCAGCTTCTCTTTGC Chr15:25710156..25710175 59.17 55
downstream ENSMUSE00000649957 Chr15:25710729..25711619 TCCTCGTAATCGTCCTGGTC Chr15:25711483..25711502 60.07 55
downstream ENSMUSE00000649953 Chr15:25712673..25712796 AGTCTCGCCGGTAGGAAAGT Chr15:25712745..25712764 60.27 55
downstream ENSMUSE00000649950 Chr15:25715558..25715801 CTCGTCCTTCAGGACACACC Chr15:25715610..25715629 60.71 60
downstream ENSMUSE00000649948 Chr15:25722906..25723001 GATGATGTCGATCCCGTTCT Chr15:25722954..25722973 59.89 50
downstream ENSMUSE00000649946 Chr15:25725438..25725531 GTCTGTGGACGAGTGGACCT Chr15:25725483..25725502 60.16 60
downstream ENSMUSE00000649943 Chr15:25726954..25727009 CAGAGGCGCATACAGAATCA Chr15:25726999..25727018 59.97 50
downstream ENSMUSE00000649940 Chr15:25729166..25729308 CAGCAGGGTTATCCAGTGGT Chr15:25729259..25729278 59.99 55
downstream ENSMUSE00000649936 Chr15:25729796..25729990 AGGGAGGACATCTTCGGACT Chr15:25729844..25729863 60.07 55
downstream ENSMUSE00000649933 Chr15:25730878..25731035 CCATACACGGTGACATTCCA Chr15:25730905..25730924 60.24 50
downstream ENSMUSE00000649931 Chr15:25733018..25733141 CTCCACCACGTCAGAGTTGA Chr15:25733050..25733069 59.86 55
downstream ENSMUSE00000649929 Chr15:25734054..25734351 GGCACCTTGTTGGTCTGTTT Chr15:25734243..25734262 60.01 50
downstream ENSMUSE00000649927 Chr15:25735266..25735473 CTTCTCCATCTCGGTTCCTG Chr15:25735305..25735324 59.8 55
downstream ENSMUSE00000649925 Chr15:25736298..25736434 CCACCTGTCCGTTGTACTCA Chr15:25736382..25736401 59.59 55
downstream ENSMUSE00000649922 Chr15:25736812..25736932 GCAAACTCCACGCTGTCTTT Chr15:25736919..25736938 60.44 50
downstream ENSMUSE00000649920 Chr15:25737491..25737943 TGATCCATAACCAGGCCACT Chr15:25737928..25737947 60.34 50
downstream ENSMUSE00000649919 Chr15:25740114..25740305 CCGCTCGTCAACTACGATCT Chr15:25740284..25740303 60.42 55
downstream ENSMUSE00000404800 Chr15:25741876..25743426 TTGTGAGCAGACTCGTTTGG Chr15:25743335..25743354 60.03 50
downstream ENSMUSE00000562965 Chr15:25741876..25741980 CACAGAGCGTGTGGTGCTAT Chr15:25741956..25741975 59.93 55
downstream ENSMUSE00000684019 Chr15:25741876..25743428 TTGTGAGCAGACTCGTTTGG Chr15:25743335..25743354 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAAGCCAAACACACAGAAGG Chr15:25673994..25674014 59.33 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCAAAGTCTTGCGTGACTGG Chr15:25674052..25674072 60.03 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022272