Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI19365
Trapped Gene
Melk (ENSMUSG00000035683)
Vector Insertion
Chr 4: 44313945 - 44316442
Public Clones not available
Private Clones OST393373 (lexicon) OST358326 (lexicon) OST182842 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000286813 (Chr4:44313822..44313944 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGTTCCGGCTTTGCTAGACT Chr4:44313854..44313873 60.01 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000286813 (Chr4:44313822..44313944 +)
Downstram Exon
ENSMUSE00000286803 (Chr4:44316443..44316538 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGTTCCGGCTTTGCTAGACT Chr4:44313854..44313873 60.01 50 TCCACCTGGAAGGTTATTCAA Chr4:44316480..44316500 59.42 42.86

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000286813 Chr4:44313822..44313944 TGTTCCGGCTTTGCTAGACT Chr4:44313854..44313873 60.01 50

*** Putative Vector Insertion (Chr 4: 44313945 - 44316442) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000286803 Chr4:44316443..44316538 TCCACCTGGAAGGTTATTCAA Chr4:44316480..44316500 59.42 42.86
downstream ENSMUSE00000286795 Chr4:44318904..44318989 CCCTAGCGCATTCTTATCCA Chr4:44318992..44319011 60.19 50
downstream ENSMUSE00000286783 Chr4:44319862..44319978 ACTCTTCAGCGCATCGATCT Chr4:44319909..44319928 60.13 50
downstream ENSMUSE00000286772 Chr4:44321778..44321921 AGGCGATCCTGGGAGATTAT Chr4:44321830..44321849 59.89 50
downstream ENSMUSE00000286762 Chr4:44323162..44323230 TTTGCACAAAGACCAAAGTCA Chr4:44323225..44323245 59.35 38.1
downstream ENSMUSE00000286755 Chr4:44325036..44325128 CATAAGCAAGGCTCCCACAG Chr4:44325087..44325106 60.79 55
downstream ENSMUSE00000714339 Chr4:44330896..44330994 CCACACATGAGCACATACAGG Chr4:44330944..44330964 60.04 52.38
downstream ENSMUSE00000719937 Chr4:44330896..44330994 CCACACATGAGCACATACAGG Chr4:44330944..44330964 60.04 52.38
downstream ENSMUSE00000286741 Chr4:44337072..44337140 TGGGAGAGAGCCACTTAGGA Chr4:44337111..44337130 59.94 55
downstream ENSMUSE00000674542 Chr4:44337072..44337140 TGGGAGAGAGCCACTTAGGA Chr4:44337111..44337130 59.94 55
downstream ENSMUSE00000674541 Chr4:44338513..44338611 CTCCACGGGACAGCTGTAAT Chr4:44338596..44338615 60.13 55
downstream ENSMUSE00000711030 Chr4:44338513..44338611 CTCCACGGGACAGCTGTAAT Chr4:44338596..44338615 60.13 55
downstream ENSMUSE00000714400 Chr4:44338513..44338611 CTCCACGGGACAGCTGTAAT Chr4:44338596..44338615 60.13 55
downstream ENSMUSE00000286730 Chr4:44345752..44345838 AAGCTCTGTCACGCAATCCT Chr4:44345790..44345809 60.02 50
downstream ENSMUSE00000674558 Chr4:44345752..44345838 AAGCTCTGTCACGCAATCCT Chr4:44345790..44345809 60.02 50
downstream ENSMUSE00000286727 Chr4:44353498..44353617 GTGGCTGTGAGGTGATCGTA Chr4:44353526..44353545 59.71 55
downstream ENSMUSE00000660800 Chr4:44353498..44353617 GTGGCTGTGAGGTGATCGTA Chr4:44353526..44353545 59.71 55
downstream ENSMUSE00000286720 Chr4:44357785..44357895 TGGCGTCTTGGGAGCTAATA Chr4:44357895..44357914 60.73 50
downstream ENSMUSE00000660799 Chr4:44357785..44357895 TGGCGTCTTGGGAGCTAATA Chr4:44357895..44357914 60.73 50
downstream ENSMUSE00000286714 Chr4:44359996..44360227 ACTGGAATCTTCGGCTCAGA Chr4:44360165..44360184 59.95 50
downstream ENSMUSE00000674557 Chr4:44359996..44360227 ACTGGAATCTTCGGCTCAGA Chr4:44360165..44360184 59.95 50
downstream ENSMUSE00000286705 Chr4:44362716..44362812 GTGTTCTAACCGGGGCTTCT Chr4:44362757..44362776 60.5 55
downstream ENSMUSE00000674555 Chr4:44362716..44362812 GTGTTCTAACCGGGGCTTCT Chr4:44362757..44362776 60.5 55
downstream ENSMUSE00000286695 Chr4:44363814..44363982 TGAGATCCACGTCCATTGAA Chr4:44363842..44363861 60.05 45
downstream ENSMUSE00000674554 Chr4:44363814..44363982 TGAGATCCACGTCCATTGAA Chr4:44363842..44363861 60.05 45
downstream ENSMUSE00000286686 Chr4:44373744..44373847 CACCAGGCGAGTTGTAGTCA Chr4:44373779..44373798 59.9 55
downstream ENSMUSE00000674552 Chr4:44373744..44373847 CACCAGGCGAGTTGTAGTCA Chr4:44373779..44373798 59.9 55
downstream ENSMUSE00000286680 Chr4:44376582..44377154 ACAGTTGCCTTGGATTCAGC Chr4:44376956..44376975 60.26 50
downstream ENSMUSE00000674546 Chr4:44376582..44377154 ACAGTTGCCTTGGATTCAGC Chr4:44376956..44376975 60.26 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCCTTCAGGTCAGTTCTTCC Chr4:44313938..44313958 58.72 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCCTTCAGGTCAGTTCTTCC Chr4:44313938..44313958 58.72 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000035683