Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6534
Trapped Gene
Dhrs7b (ENSMUSG00000042569)
Vector Insertion
Chr 11: 60657846 - 60662602
Public Clones CSE042 (baygenomics) CMHD-GT_486F4-3 (cmhd)
Private Clones OST445833 (lexicon) OST213656 (lexicon)
Severity of mutation (?) Insertion after 17% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000242395 (Chr11:60657667..60657845 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CGCTCAAAGGCTTACCTGAG Chr11:60657779..60657798 60.15 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000242395 (Chr11:60657667..60657845 +)
Downstram Exon
ENSMUSE00000242388 (Chr11:60662603..60662706 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CGCTCAAAGGCTTACCTGAG Chr11:60657779..60657798 60.15 55 TCACATTTCGTCCACAGAGG Chr11:60662665..60662684 59.68 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000677766 Chr11:60644181..60644233 GACCATGATCTCTCCGTCCT Chr11:60644210..60644229 59.06 55
upstream ENSMUSE00000242398 Chr11:60644189..60644233 GACCATGATCTCTCCGTCCT Chr11:60644210..60644229 59.06 55
upstream ENSMUSE00000242395 Chr11:60657667..60657845 CGCTCAAAGGCTTACCTGAG Chr11:60657779..60657798 60.15 55
upstream ENSMUSE00000677765 Chr11:60657667..60657845 CGCTCAAAGGCTTACCTGAG Chr11:60657779..60657798 60.15 55

*** Putative Vector Insertion (Chr 11: 60657846 - 60662602) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000242388 Chr11:60662603..60662706 TCACATTTCGTCCACAGAGG Chr11:60662665..60662684 59.68 50
downstream ENSMUSE00000242379 Chr11:60665244..60665460 GAGCAACAGGGCCAAAGTAG Chr11:60665454..60665473 59.88 55
downstream ENSMUSE00000242373 Chr11:60665921..60666013 GATCGGAAAGGAATGCTGAT Chr11:60666011..60666030 59.08 45
downstream ENSMUSE00000242367 Chr11:60669202..60669354 TCGGCAGTGACAGCATTTAC Chr11:60669340..60669359 59.87 50
downstream ENSMUSE00000391252 Chr11:60671220..60671647 CATGGATGGCACAAAGTCTG Chr11:60671338..60671357 60.11 50
downstream ENSMUSE00000677764 Chr11:60671220..60673697 CCAGTGCACCTCCTATGGTT Chr11:60672620..60672639 59.99 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATAATCGCCTTGCAGCACAT Chr11:60660896..60660916 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACACATTCAATGGCCTGCTT Chr11:60660855..60660875 60.53 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000042569