Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI8949
Trapped Gene
Pvrl2 (ENSMUSG00000062300)
Vector Insertion
Chr 7: 20304846 - 20306701
Public Clones RRE206 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 100% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000198359 (Chr7:20306702..20306855 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TATTATCGCTGCCATCATCG Chr7:20306787..20306806 59.64 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000198359 (Chr7:20306702..20306855 -)
Downstram Exon
ENSMUSE00000198354 (Chr7:20304782..20304845 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TATTATCGCTGCCATCATCG Chr7:20306787..20306806 59.64 45 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000536242 Chr7:20334564..20334818 CCGATCGGTACACGAAGAGT Chr7:20334700..20334719 60.13 55
upstream ENSMUSE00000357931 Chr7:20323361..20323723 GGCAATTACACCTGCGAGTT Chr7:20323414..20323433 60.14 50
upstream ENSMUSE00000198358 Chr7:20318264..20318560 CCGAATCACCTGGATCTCAT Chr7:20318444..20318463 59.89 50
upstream ENSMUSE00000198353 Chr7:20315961..20316078 ATCCATCTCCGGCTATGATG Chr7:20316046..20316065 59.88 50
upstream ENSMUSE00000198361 Chr7:20315387..20315535 TCTGTGGATCGAATGGTCAA Chr7:20315458..20315477 60.05 45
upstream ENSMUSE00000676814 Chr7:20309830..20310218 TCATGCTACCTCCACACGAG Chr7:20309899..20309918 59.85 55
upstream ENSMUSE00000198359 Chr7:20306702..20306855 TATTATCGCTGCCATCATCG Chr7:20306787..20306806 59.64 45

*** Putative Vector Insertion (Chr 7: 20304846 - 20306701) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000198354 Chr7:20304782..20304845 No primer for this exon
downstream ENSMUSE00000198357 Chr7:20304545..20304631 CCCCAAGGTGAAGAGTTGAG Chr7:20304586..20304605 59.69 55
downstream ENSMUSE00000485480 Chr7:20302015..20303136 CAGATGCCCAGATTCTCCAT Chr7:20302047..20302066 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCGGATGAGGAAGAAGAGTG Chr7:20306697..20306717 59.95 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCGGATGAGGAAGAAGAGTG Chr7:20306697..20306717 59.95 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000062300