Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24707
Trapped Gene
Paip2b (ENSMUSG00000045896)
Vector Insertion
Chr 6: 83764864 - 83781264
Public Clones (sanger) (sanger)
Private Clones OST209344 (lexicon) OST145066 (lexicon)
Severity of mutation (?) Insertion after 7% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000559040 (Chr6:83781265..83781322 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GATGACGACGCTGAACACAG Chr6:83781275..83781294 60.47 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000559040 (Chr6:83781265..83781322 -)
Downstram Exon
ENSMUSE00000390940 (Chr6:83764716..83764863 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GATGACGACGCTGAACACAG Chr6:83781275..83781294 60.47 55 TTCTGCAAATGGGTTTTCCT Chr6:83764733..83764752 59.55 40

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000653661 Chr6:83781629..83781735 AGGTACCTCGAGCGTTTGTG Chr6:83781705..83781724 60.31 55
upstream ENSMUSE00000559040 Chr6:83781265..83781322 GATGACGACGCTGAACACAG Chr6:83781275..83781294 60.47 55

*** Putative Vector Insertion (Chr 6: 83764864 - 83781264) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000390940 Chr6:83764716..83764863 TTCTGCAAATGGGTTTTCCT Chr6:83764733..83764752 59.55 40
downstream ENSMUSE00000350403 Chr6:83759893..83760069 GTCCTGGTCCTCTTCATCCA Chr6:83759973..83759992 60.05 55
downstream ENSMUSE00000615086 Chr6:83756056..83758886 CGGGTACAATTGTGCTTCCT Chr6:83756571..83756590 59.99 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr6:83769193..83769213 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGAATGATCGTGACTGGGAAA Chr6:83775200..83775221 59.54 42.86 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 AACAGCCAGTAATCGCCTTG Chr6:83775261..83775281 60.27 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 AAGAATGCCAATTTGGGAAC Chr6:83775308..83775328 58.88 40 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000045896