Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI17818
Trapped Gene
Tmtc1 (ENSMUSG00000030306)
Vector Insertion
Chr 6: 148243205 - 148254519
Public Clones IST12288A10 (tigm) IST11517A1 (tigm) IST12803G10 (tigm) IST12415B10 (tigm)
IST10206F2 (tigm) IST14118C6 (tigm) IST10226F1 (tigm) IST12090D4 (tigm)
Private Clones OST435961 (lexicon) OST434921 (lexicon)
Severity of mutation (?) Insertion after 51% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000197285 (Chr6:148254520..148254641 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGGCCGAGAGAGTCCTGTA Chr6:148254529..148254548 60.41 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000197285 (Chr6:148254520..148254641 -)
Downstram Exon
ENSMUSE00000275629 (Chr6:148243037..148243204 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGGCCGAGAGAGTCCTGTA Chr6:148254529..148254548 60.41 60 GCCCAGCACAGAGCTTACTC Chr6:148243130..148243149 60.16 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000650123 Chr6:148392423..148392845 CTAACGACTTCTGGGGCAAG Chr6:148392482..148392501 59.87 55
upstream ENSMUSE00000650122 Chr6:148374329..148374506 GTCTTCAAGAATCGCGGACT Chr6:148374381..148374400 59.43 50
upstream ENSMUSE00000650120 Chr6:148364232..148364305 CTAGCATGCCTCCTGTTCCT Chr6:148364256..148364275 59.45 55
upstream ENSMUSE00000650118 Chr6:148361164..148361340 ATGCTGGTGAAGGAGACAGG Chr6:148361230..148361249 60.26 55
upstream ENSMUSE00000650116 Chr6:148359560..148359763 GCTTCCCACACAAAGACTCC Chr6:148359646..148359665 59.7 55
upstream ENSMUSE00000650115 Chr6:148284773..148284854 GTTTTTAAAGCGGGCCATTT Chr6:148284787..148284806 60.31 40
upstream ENSMUSE00000197286 Chr6:148284165..148284268 ATTTCCGCCTGTGGATAATG Chr6:148284233..148284252 59.78 45
upstream ENSMUSE00000197278 Chr6:148273639..148273828 TGGGAAGTATCCCTCTGGTG Chr6:148273729..148273748 59.92 55
upstream ENSMUSE00000197285 Chr6:148254520..148254641 GTGGCCGAGAGAGTCCTGTA Chr6:148254529..148254548 60.41 60

*** Putative Vector Insertion (Chr 6: 148243205 - 148254519) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000275629 Chr6:148243037..148243204 GCCCAGCACAGAGCTTACTC Chr6:148243130..148243149 60.16 60
downstream ENSMUSE00000650109 Chr6:148233379..148233492 AGGGCGGTTCTGTAGTGGTA Chr6:148233360..148233379 59.62 55
downstream ENSMUSE00000197282 Chr6:148219898..148220041 TTCCCCAGATTGAAAAGTGC Chr6:148219885..148219904 60.05 45
downstream ENSMUSE00000197302 Chr6:148211305..148211413 GCCAAGCTTGAATAGGCATC Chr6:148211302..148211321 59.81 50
downstream ENSMUSE00000197292 Chr6:148198010..148198112 TTCCAGCCTGGTAAATGTCC Chr6:148198051..148198070 59.93 50
downstream ENSMUSE00000275608 Chr6:148195925..148196060 GGTAGTGAGCCACAGCCTTC Chr6:148196009..148196028 59.87 60
downstream ENSMUSE00000275601 Chr6:148195256..148195400 AGCTTCCCTGTACACCTCCA Chr6:148195276..148195295 59.72 55
downstream ENSMUSE00000197288 Chr6:148194233..148194370 TTTGGTCTGACCCATCACTG Chr6:148194316..148194335 59.52 50
downstream ENSMUSE00000197296 Chr6:148193206..148193328 TCAAAAGCTTTGTCCAGAAGG Chr6:148193185..148193205 59.48 42.86
downstream ENSMUSE00000197284 Chr6:148191710..148191787 GTCAGGATCCAGGGTCACAG Chr6:148191733..148191752 60.53 60
downstream ENSMUSE00000379302 Chr6:148185119..148186456 AGTGATAATCTGGGCGATGC Chr6:148185771..148185790 60.07 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AAACAGTTAATCGCCTTGCAG Chr6:148254454..148254475 59.42 42.86 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAAACAGTCGTGACTGGGAAA Chr6:148245455..148245476 60.14 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CAAAGCTAATCGCCTTGCAG Chr6:148245577..148245597 61.03 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 AAATCGTGACTGGGAAAACC Chr6:148254575..148254595 58.89 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000030306