Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI1110
Trapped Gene
Nasp (ENSMUSG00000028693)
Vector Insertion
Chr 4: 116299936 - 116300035
Public Clones CC0815 (sanger) CC0209 (sanger) D014G07 (ggtc) W171F03 (ggtc) H003F08 (ggtc)
P134B06 (ggtc) P102B04 (ggtc) P137H08 (ggtc) H008G08 (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
ENSMUSE00000181400 (Chr4:116299937..116300034 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CGATGGCCACAGAGTCTACA Chr4:116299978..116299997 59.85 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000181400 (Chr4:116299937..116300034 -)
Downstram Exon
ENSMUSE00000525389 (Chr4:116299937..116300083 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CGATGGCCACAGAGTCTACA Chr4:116299978..116299997 59.85 55 AGAGAATTCCGGACAGCAGA Chr4:116300027..116300046 59.95 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

*** Putative Vector Insertion (Chr 4: 116299936 - 116300035) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000181395 Chr4:116295372..116295419 TCCATTTTATCTGCCGAGGT Chr4:116295353..116295372 59.53 45
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 GGAACGATGGCCACAGTAAT Chr4:116299980..116300000 59.82 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTTTTCTTCGCAGGCAGTGT Chr4:116300026..116300046 60.44 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028693