Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI16319
Trapped Gene
Mrps27 (ENSMUSG00000041632)
Vector Insertion
Chr 13: 100182321 - 100184693
Public Clones IST12497C6 (tigm)
Private Clones OST463246 (lexicon)
Severity of mutation (?) Insertion after 81% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000294446 (Chr13:100182150..100182320 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATGGAGTCCTGAAGGTGGTG Chr13:100182163..100182182 59.96 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000294446 (Chr13:100182150..100182320 +)
Downstram Exon
ENSMUSE00000374496 (Chr13:100184694..100185517 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATGGAGTCCTGAAGGTGGTG Chr13:100182163..100182182 59.96 55 CTGTCCTGGCTGAACACTGA Chr13:100184972..100184991 60.02 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000294499 Chr13:100114745..100114830 No primer for this exon
upstream ENSMUSE00000294492 Chr13:100118287..100118364 AAAAGATGCCTGCTTTCTGC Chr13:100118289..100118308 59.6 45
upstream ENSMUSE00000294486 Chr13:100132920..100132990 TGCATCCTTAATGGACAAAGC Chr13:100132927..100132947 60.09 42.86
upstream ENSMUSE00000294479 Chr13:100135180..100135238 ACATTGCATCCCGAGAAGAT Chr13:100135190..100135209 59.51 45
upstream ENSMUSE00000294474 Chr13:100170217..100170331 GAGACTGGACCATCCACAGC Chr13:100170249..100170268 60.69 60
upstream ENSMUSE00000294469 Chr13:100173459..100173537 No primer for this exon
upstream ENSMUSE00000294465 Chr13:100174943..100175058 ACCGTTGCCTAGCAGAGAAG Chr13:100175027..100175046 59.64 55
upstream ENSMUSE00000407388 Chr13:100179764..100179866 CTGGCCTGAAACAAAGGAAC Chr13:100179804..100179823 59.71 50
upstream ENSMUSE00000433773 Chr13:100181233..100181375 ACACCAGGCTACCTCGACAG Chr13:100181292..100181311 60.32 60
upstream ENSMUSE00000294446 Chr13:100182150..100182320 ATGGAGTCCTGAAGGTGGTG Chr13:100182163..100182182 59.96 55

*** Putative Vector Insertion (Chr 13: 100182321 - 100184693) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000374496 Chr13:100184694..100185517 CTGTCCTGGCTGAACACTGA Chr13:100184972..100184991 60.02 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGAACCAGAACAGTCCAAGC Chr13:100182276..100182296 59.7 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGAACCAGAACAGTCCAAGC Chr13:100182276..100182296 59.7 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000041632