Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI11645
Trapped Gene
Nasp (ENSMUSG00000028693)
Vector Insertion
Chr 4: 116291586 - 116295371
Public Clones (sanger) P078D09 (ggtc) H003F08 (ggtc) PST23465-NR (escells) PST19125-NR (escells)
Private Clones OST351458 (lexicon) OST209923 (lexicon) OST68145 (lexicon) OST67988 (lexicon)
OST13724 (lexicon)
Severity of mutation (?) Insertion after 5% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000181395 (Chr4:116295372..116295419 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCTCGGCAGATAAAATGGAG Chr4:116295374..116295393 59.66 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000181395 (Chr4:116295372..116295419 -)
Downstram Exon
ENSMUSE00000181391 (Chr4:116291475..116291585 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCTCGGCAGATAAAATGGAG Chr4:116295374..116295393 59.66 50 TGTCACCCATCACCAGATGT Chr4:116291496..116291515 59.8 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000407904 Chr4:116300495..116300556 GCGTTTCCATCCGTTAAGAC Chr4:116300530..116300549 59.57 50
upstream ENSMUSE00000181400 Chr4:116299937..116300034 CGATGGCCACAGAGTCTACA Chr4:116299978..116299997 59.85 55
upstream ENSMUSE00000505449 Chr4:116299937..116300102 TCTGCTGTCCGGAATTCTCT Chr4:116300049..116300068 59.95 50
upstream ENSMUSE00000525389 Chr4:116299937..116300083 TCTGCTGTCCGGAATTCTCT Chr4:116300049..116300068 59.95 50
upstream ENSMUSE00000181395 Chr4:116295372..116295419 CCTCGGCAGATAAAATGGAG Chr4:116295374..116295393 59.66 50

*** Putative Vector Insertion (Chr 4: 116291586 - 116295371) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000181391 Chr4:116291475..116291585 TGTCACCCATCACCAGATGT Chr4:116291496..116291515 59.8 50
downstream ENSMUSE00000181404 Chr4:116286935..116287015 TCCACACTCATTAGCCGTTTC Chr4:116286957..116286977 60.12 47.62
downstream ENSMUSE00000181402 Chr4:116284577..116284686 CCCAACACTCCATTCTCCAT Chr4:116284644..116284663 59.78 50
downstream ENSMUSE00000470333 Chr4:116283008..116283982 CATCCGCTGACTTGACTTGA Chr4:116283451..116283470 59.98 50
downstream ENSMUSE00000259672 Chr4:116278289..116278368 CCTCTTCCTCTGAGCCTTCA Chr4:116278323..116278342 59.67 55
downstream ENSMUSE00000259667 Chr4:116277514..116277599 CAGCATATCCCAGGCAAGTT Chr4:116277518..116277537 60.1 50
downstream ENSMUSE00000259660 Chr4:116277346..116277419 CAGCATAAAGCTGGGCTTCT Chr4:116277366..116277385 59.62 50
downstream ENSMUSE00000259655 Chr4:116276768..116276956 ACGATCGTGAGCTTCCAAAT Chr4:116276858..116276877 59.7 45
downstream ENSMUSE00000181393 Chr4:116276500..116276666 AGTGCCAGTTCGGCTACATT Chr4:116276491..116276510 59.76 50
downstream ENSMUSE00000181399 Chr4:116275493..116275549 GGATACTGAGGCACCAGCTC Chr4:116275474..116275493 59.83 60
downstream ENSMUSE00000181403 Chr4:116275331..116275408 CGCCATCTGTTGGTTTTCTA Chr4:116275359..116275378 58.77 45
downstream ENSMUSE00000181396 Chr4:116274697..116274825 CACTGCCTCCATTCACCTCT Chr4:116274725..116274744 60.26 55
downstream ENSMUSE00000631434 Chr4:116274214..116274398 TTGGCTCTCAGCCTGATTCT Chr4:116274353..116274372 60.1 50
downstream ENSMUSE00000475846 Chr4:116273657..116274398 TTTAGCAGACCCCATCCAAG Chr4:116273945..116273964 60.07 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCTGTGCATGCTAGGAAAGC Chr4:116292345..116292365 59.18 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCTGTGCATGCTAGGAAAGC Chr4:116292345..116292365 59.18 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CCTCGGCAGATAAAATGGAG Chr4:116292372..116292392 59.66 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CTGATCTACGTGACTGGGAAAA Chr4:116292355..116292377 59.24 45.46 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028693