Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI21519
Trapped Gene
Lsm4 (ENSMUSG00000031848)
Vector Insertion
Chr 8: 73202034 - 73202258
Public Clones CMHD-GT_390A11-3 (cmhd) IST11817D7 (tigm)
Private Clones OST327360 (lexicon) OST279074 (lexicon) OST247515 (lexicon) OST21624 (lexicon)
Severity of mutation (?) Insertion after 78% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000213764 (Chr8:73201853..73202033 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGGTGACAAGTTCTGGAGGA Chr8:73201855..73201874 59.23 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000213764 (Chr8:73201853..73202033 +)
Downstram Exon
ENSMUSE00000636306 (Chr8:73202259..73202642 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGGTGACAAGTTCTGGAGGA Chr8:73201855..73201874 59.23 50 AAGTGGGGCTTCACACAAAG Chr8:73202424..73202443 60.15 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000682766 Chr8:73197278..73197356 TTATCGCTGCAGGGAAGATG Chr8:73197337..73197356 61.27 50
upstream ENSMUSE00000225998 Chr8:73200454..73200495 TGAAGACAGCGCAGAATCAC Chr8:73200470..73200489 60.14 50
upstream ENSMUSE00000213763 Chr8:73201677..73201775 GAATGGGGAGACCTACAACG Chr8:73201691..73201710 59.4 55
upstream ENSMUSE00000213764 Chr8:73201853..73202033 TGGTGACAAGTTCTGGAGGA Chr8:73201855..73201874 59.23 50

*** Putative Vector Insertion (Chr 8: 73202034 - 73202258) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000636306 Chr8:73202259..73202642 AAGTGGGGCTTCACACAAAG Chr8:73202424..73202443 60.15 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTCCCTGTGAGGATGAGGTG Chr8:73202040..73202060 60.53 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTCCCTGTGAGGATGAGGTG Chr8:73202040..73202060 60.53 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031848