Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20484
Trapped Gene
Creb3l1 (ENSMUSG00000027230)
Vector Insertion
Chr 2: 91842231 - 91864160
Public Clones IST13151D3 (tigm) IST13298H11 (tigm) IST13151D3 (tigm)
Private Clones OST360894 (lexicon) OST78997 (lexicon)
Severity of mutation (?) Insertion after 6% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000357520 (Chr2:91864161..91864327 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGGACTTGGGAGACCTGAA Chr2:91864186..91864205 60.23 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000357520 (Chr2:91864161..91864327 -)
Downstram Exon
ENSMUSE00000167183 (Chr2:91842002..91842230 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGGACTTGGGAGACCTGAA Chr2:91864186..91864205 60.23 55 CTCCATCTTGACAGGCACAA Chr2:91841993..91842012 59.83 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000357520 Chr2:91864161..91864327 CTGGACTTGGGAGACCTGAA Chr2:91864186..91864205 60.23 55

*** Putative Vector Insertion (Chr 2: 91842231 - 91864160) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000167183 Chr2:91842002..91842230 CTCCATCTTGACAGGCACAA Chr2:91841993..91842012 59.83 50
downstream ENSMUSE00000167189 Chr2:91835414..91835598 TGATGGAGCACAGCTTGTTT Chr2:91835529..91835548 59.44 45
downstream ENSMUSE00000167186 Chr2:91833428..91833506 GGCTCTGCTTTGATCACTGG Chr2:91833441..91833460 60.94 55
downstream ENSMUSE00000167193 Chr2:91832044..91832201 No primer for this exon
downstream ENSMUSE00000167188 Chr2:91831283..91831432 AAGGTCCGCTTCTCCTCTTC Chr2:91831355..91831374 59.96 55
downstream ENSMUSE00000167192 Chr2:91831066..91831124 No primer for this exon
downstream ENSMUSE00000167191 Chr2:91830864..91830932 TGGCAGTCTCTAGGGTTTCC Chr2:91830846..91830865 59.28 55
downstream ENSMUSE00000360669 Chr2:91827193..91827292 GCTGCCATCTTGTACGGTCT Chr2:91827196..91827215 60.28 55
downstream ENSMUSE00000299441 Chr2:91826318..91826444 AGTCATTGAGCCGGAAGAGA Chr2:91826345..91826364 59.95 50
downstream ENSMUSE00000370902 Chr2:91823322..91823589 GTGCCGTTGTCATCAGTGTC Chr2:91823342..91823361 60.17 55
downstream ENSMUSE00000502278 Chr2:91822487..91823164 TAGCAAAGCCCGCACTAACT Chr2:91822623..91822642 60.04 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCCAAGGGTCCTACACCTTC Chr2:91858186..91858206 60.74 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCCAAGGGTCCTACACCTTC Chr2:91858186..91858206 60.74 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CTGATGGTGAAGGCTCCATT Chr2:91858335..91858355 60.07 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CTGATGGTGAAGGCTCCATT Chr2:91858335..91858355 60.07 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027230