Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14439
Trapped Gene
Slc25a12 (ENSMUSG00000027010)
Vector Insertion
Chr 2: 71204583 - 71205641
Public Clones FHCRC-GT-S12-8F1 (fhcrc)
Private Clones OST378503 (lexicon)
Severity of mutation (?) Insertion after 1% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000662334 (Chr2:71205570..71205640 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000662334 (Chr2:71205570..71205640 -)
Downstram Exon
ENSMUSE00000662333 (Chr2:71204584..71204637 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon AGGATCACCTCGTTTGGTTG Chr2:71204589..71204608 59.97 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000662334 Chr2:71205570..71205640 No primer for this exon
upstream ENSMUSE00000662333 Chr2:71204584..71204637 CAACCAAACGAGGTGATCCT Chr2:71204611..71204630 59.97 50

*** Putative Vector Insertion (Chr 2: 71204583 - 71205641) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000662332 Chr2:71181947..71182089 GGATCGTTGTACAGGCCAAG Chr2:71181985..71182004 60.52 55
downstream ENSMUSE00000320612 Chr2:71171662..71171777 TCTCCATTTCCGCTCTTGTC Chr2:71171651..71171670 60.34 50
downstream ENSMUSE00000165429 Chr2:71162033..71162172 TCTGCCCAAAAATCTCCTTG Chr2:71162124..71162143 60.18 45
downstream ENSMUSE00000165435 Chr2:71153040..71153186 GAAATCCAGACCCGAGATCA Chr2:71153084..71153103 60.01 50
downstream ENSMUSE00000566456 Chr2:71150482..71150620 TATCTTTCCTCGTGCCTGCT Chr2:71150475..71150494 59.98 50
downstream ENSMUSE00000165430 Chr2:71149520..71149613 No primer for this exon
downstream ENSMUSE00000320519 Chr2:71146113..71146197 GCGGAGCTATTCTCTCGATG Chr2:71146138..71146157 60.08 55
downstream ENSMUSE00000320512 Chr2:71136627..71136708 CTGGAGCCAGATAGGTCTGC Chr2:71136651..71136670 59.97 60
downstream ENSMUSE00000566451 Chr2:71134722..71134880 TCCAAAGAAACCCTCGTAGC Chr2:71134710..71134729 59.31 50
downstream ENSMUSE00000165434 Chr2:71131729..71131781 TTTCTGGTGCAACCCCTATC Chr2:71131724..71131743 59.93 50
downstream ENSMUSE00000165424 Chr2:71131417..71131497 GCAACCTCCAGCAAGGATTT Chr2:71131395..71131414 61.51 50
downstream ENSMUSE00000566446 Chr2:71120527..71120667 CCTGAAGCACATTCAGAGCA Chr2:71120531..71120550 60.14 50
downstream ENSMUSE00000165433 Chr2:71117573..71117711 GTGAGCGTACACGGGAAAAT Chr2:71117624..71117643 60 50
downstream ENSMUSE00000165428 Chr2:71114698..71114856 CCTTCTTCCCGGAGAATCTT Chr2:71114705..71114724 59.65 50
downstream ENSMUSE00000566445 Chr2:71114489..71114579 GAGGTGAGGATCGAAACACC Chr2:71114534..71114553 59.51 55
downstream ENSMUSE00000662331 Chr2:71112803..71113410 CCTGGGCGTATGATCACTTT Chr2:71113052..71113071 59.96 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCTTCCGTGACTGGGAAAAC Chr2:71205575..71205595 61.96 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 2 TGCTGCTGCTTTTATTGCAT Chr2:71205533..71205553 59.61 40 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 3 TGCTGCTGCTTTTATTGCAT Chr2:71205533..71205553 59.61 40 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027010