Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI27403
Trapped Gene
Tmem161b (ENSMUSG00000035762)
Vector Insertion
Chr 13: 84411955 - 84422315
Public Clones not available
Private Clones OST60572 (lexicon)
Severity of mutation (?) Insertion after 31% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000470322 (Chr13:84411776..84411954 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AATACCAGTGGCTGGTGGAC Chr13:84411794..84411813 59.85 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000470322 (Chr13:84411776..84411954 +)
Downstram Exon
ENSMUSE00000248330 (Chr13:84422316..84422467 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AATACCAGTGGCTGGTGGAC Chr13:84411794..84411813 59.85 55 TGCCATTGCTTTAACGAAGA Chr13:84422424..84422443 59.44 40

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000708985 Chr13:84361931..84362039 GTTTCGAGAGCTCCGTTTTG Chr13:84361943..84361962 59.99 50
upstream ENSMUSE00000713302 Chr13:84361931..84362039 GTTTCGAGAGCTCCGTTTTG Chr13:84361943..84361962 59.99 50
upstream ENSMUSE00000400401 Chr13:84390835..84390938 CTTGCTCGATGGCTACTCTG Chr13:84390910..84390929 58.78 55
upstream ENSMUSE00000403723 Chr13:84397366..84397449 TCAACATCCTTCCGAAGAAGA Chr13:84397378..84397398 59.8 42.86
upstream ENSMUSE00000363602 Chr13:84399748..84399845 AGCCACTAACCATTCCAAAAGA Chr13:84399774..84399795 60 40.91
upstream ENSMUSE00000470322 Chr13:84411776..84411954 AATACCAGTGGCTGGTGGAC Chr13:84411794..84411813 59.85 55
upstream ENSMUSE00000519398 Chr13:84411776..84411932 AATACCAGTGGCTGGTGGAC Chr13:84411794..84411813 59.85 55

*** Putative Vector Insertion (Chr 13: 84411955 - 84422315) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000248330 Chr13:84422316..84422467 TGCCATTGCTTTAACGAAGA Chr13:84422424..84422443 59.44 40
downstream ENSMUSE00000248327 Chr13:84422909..84422969 TGCATTGCACTGTCTGAAAA Chr13:84422942..84422961 58.99 40
downstream ENSMUSE00000248321 Chr13:84423590..84423730 AAGCCCCAATGAGTGAACAG Chr13:84423657..84423676 60.11 50
downstream ENSMUSE00000248317 Chr13:84426290..84426403 ATTGGTTTCACCCAGAGCAG Chr13:84426355..84426374 60.11 50
downstream ENSMUSE00000248311 Chr13:84431984..84432158 TATGCTTGCAGGTGACTTCG Chr13:84432082..84432101 60.01 50
downstream ENSMUSE00000518452 Chr13:84432986..84433082 CATACTGCAGGGCAATGACA Chr13:84433034..84433053 60.69 50
downstream ENSMUSE00000640906 Chr13:84434208..84435569 AAGGCGGCTTCTGGATAAAT Chr13:84434247..84434266 60.06 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTCAGATAATCGCCTTGCAG Chr13:84412000..84412020 59.02 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGCCTCCTGGTTTTGTCTT Chr13:84414907..84414927 59.19 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000035762