Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20299
Trapped Gene
2900011O08Rik (ENSMUSG00000044117)
Vector Insertion
Chr 16: 14049600 - 14089038
Public Clones not available
Private Clones OST367866 (lexicon) OST357105 (lexicon) OST352254 (lexicon) OST300487 (lexicon)
OST187058 (lexicon) OST185744 (lexicon) OST142019 (lexicon) OST121565 (lexicon)
OST109489 (lexicon) OST62656 (lexicon) OST47184 (lexicon) OST47178 (lexicon)
OST46190 (lexicon) OST44590 (lexicon) OST38860 (lexicon) OST38859 (lexicon)
OST32853 (lexicon)
Severity of mutation (?) Insertion after 67% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000358544 (Chr16:14049476..14049599 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGTCCGTCGAGAGTCAGAAC Chr16:14049570..14049589 59.84 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000358544 (Chr16:14049476..14049599 +)
Downstram Exon
ENSMUSE00000399792 (Chr16:14089039..14089112 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGTCCGTCGAGAGTCAGAAC Chr16:14049570..14049589 59.84 60 TGCTTCAGGTCCATGATGTC Chr16:14089086..14089105 59.64 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000378060 Chr16:13986721..13986986 GAAGCCTCTCAGGCGTTATG Chr16:13986925..13986944 59.98 55
upstream ENSMUSE00000717969 Chr16:13986750..13986986 GAAGCCTCTCAGGCGTTATG Chr16:13986925..13986944 59.98 55
upstream ENSMUSE00000358544 Chr16:14049476..14049599 GGTCCGTCGAGAGTCAGAAC Chr16:14049570..14049589 59.84 60

*** Putative Vector Insertion (Chr 16: 14049600 - 14089038) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000399792 Chr16:14089039..14089112 TGCTTCAGGTCCATGATGTC Chr16:14089086..14089105 59.64 50
downstream ENSMUSE00000358904 Chr16:14093982..14094096 GCATCATCCACCAGGAAGTC Chr16:14094081..14094100 60.48 55
downstream ENSMUSE00000414187 Chr16:14095997..14096079 GGAAATCAGCCATTTCCTTG Chr16:14096034..14096053 59.51 45
downstream ENSMUSE00000715476 Chr16:14095997..14096079 GGAAATCAGCCATTTCCTTG Chr16:14096034..14096053 59.51 45
downstream ENSMUSE00000392859 Chr16:14099886..14101593 CTCTGCTACCTGGTCCAAGC Chr16:14100726..14100745 60.01 60
downstream ENSMUSE00000720192 Chr16:14099886..14100321 TGTTTTAAGGGAGGGGCTTT Chr16:14100283..14100302 59.94 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGACTTAGCTAATCGCCTTGC Chr16:14073642..14073663 60.37 52.38 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTTAGCCGTGACTGGGAAAA Chr16:14073645..14073665 60.24 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000044117