Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI26348
Trapped Gene
Slc25a12 (ENSMUSG00000027010)
Vector Insertion
Chr 2: 71120668 - 71131416
Public Clones IST10323B6 (tigm) IST12528F9 (tigm) IST10233F4 (tigm) IST12677A7 (tigm)
IST10323B6 (tigm)
Private Clones OST123999 (lexicon) OST7421 (lexicon)
Severity of mutation (?) Insertion after 64% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000165424 (Chr2:71131417..71131497 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTCCGGGACAAATTTACCAA Chr2:71131466..71131485 59.66 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000165424 (Chr2:71131417..71131497 -)
Downstram Exon
ENSMUSE00000566446 (Chr2:71120527..71120667 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTCCGGGACAAATTTACCAA Chr2:71131466..71131485 59.66 45 CCTGAAGCACATTCAGAGCA Chr2:71120531..71120550 60.14 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
upstream ENSMUSE00000662332 Chr2:71181947..71182089 ACCCAAAGATTGTGCAGCTC Chr2:71181979..71181998 60.26 50
upstream ENSMUSE00000320612 Chr2:71171662..71171777 GACAAGAGCGGAAATGGAGA Chr2:71171673..71171692 60.34 50
upstream ENSMUSE00000165429 Chr2:71162033..71162172 CAAGGAGATTTTTGGGCAGA Chr2:71162146..71162165 60.18 45
upstream ENSMUSE00000165435 Chr2:71153040..71153186 GATCTCGGGTCTGGATTTCA Chr2:71153105..71153124 60.01 50
upstream ENSMUSE00000566456 Chr2:71150482..71150620 AGCAGGCACGAGGAAAGATA Chr2:71150497..71150516 59.98 50
upstream ENSMUSE00000165430 Chr2:71149520..71149613 GCCATACGCTATGGACAGGT Chr2:71149577..71149596 59.99 55
upstream ENSMUSE00000320519 Chr2:71146113..71146197 TGACTCTGGCAGACATCGAG Chr2:71146173..71146192 60.14 55
upstream ENSMUSE00000320512 Chr2:71136627..71136708 No primer for this exon
upstream ENSMUSE00000566451 Chr2:71134722..71134880 CTGCGGTGTACCCCATAGAC Chr2:71134849..71134868 60.4 60
upstream ENSMUSE00000165434 Chr2:71131729..71131781 GATAGGGGTTGCACCAGAAA Chr2:71131746..71131765 59.93 50
upstream ENSMUSE00000165424 Chr2:71131417..71131497 GTCCGGGACAAATTTACCAA Chr2:71131466..71131485 59.66 45

*** Putative Vector Insertion (Chr 2: 71120668 - 71131416) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
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 TCTTCCCAGCAAAGGGATAA Chr2:71122362..71122382 59.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCTCACGTGACTGGGAAAAC Chr2:71122350..71122370 60.54 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 TGCTCTTCTAATCGCCTTGC Chr2:71122435..71122455 60.63 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CCGGGACAAATTTACCAAAA Chr2:71122462..71122482 59.67 40 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027010