Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI5688
Trapped Gene
Nasp (ENSMUSG00000028693)
Vector Insertion
Chr 4: 116278369 - 116283007
Public Clones CSI625 (baygenomics) H010H05 (ggtc) H006F04 (ggtc) PST12735-NL (escells)
PST7730-NR (escells) PST15407-NL (escells) PST11659-NL (escells) PST8586-NR (escells)
PST14790-NR (escells) PST11440-NR (escells) PST443-1 (escells)
Private Clones not available
Severity of mutation (?) Insertion after 60% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000470333 (Chr4:116283008..116283982 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TCAAGTCAAGTCAGCGGATG Chr4:116283473..116283492 59.98 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000470333 (Chr4:116283008..116283982 -)
Downstram Exon
ENSMUSE00000259672 (Chr4:116278289..116278368 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TCAAGTCAAGTCAGCGGATG Chr4:116283473..116283492 59.98 50 CCTCTTCCTCTGAGCCTTCA Chr4:116278323..116278342 59.67 55

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
upstream ENSMUSE00000181391 Chr4:116291475..116291585 ACATCTGGTGATGGGTGACA Chr4:116291518..116291537 59.8 50
upstream ENSMUSE00000181404 Chr4:116286935..116287015 GAAACGGCTAATGAGTGTGGA Chr4:116286979..116286999 60.12 47.62
upstream ENSMUSE00000181402 Chr4:116284577..116284686 TGCCTTAGAAGGAGTGCATGT Chr4:116284642..116284662 59.89 47.62
upstream ENSMUSE00000470333 Chr4:116283008..116283982 TCAAGTCAAGTCAGCGGATG Chr4:116283473..116283492 59.98 50

*** Putative Vector Insertion (Chr 4: 116278369 - 116283007) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
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 TGCAGTGGGTGGAGTACATT Chr4:116282965..116282985 59.01 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGCAGTGGGTGGAGTACATT Chr4:116282965..116282985 59.01 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 AGATAATCGCCTTGCAGCAC Chr4:116280915..116280935 60.38 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 GATTCGTGACTGGGAAAACC Chr4:116283916..116283936 59.39 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028693