Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32122
Trapped Gene
Tshz2 (ENSMUSG00000047907)
Vector Insertion
Chr 2: 169459280 - 169616465
Public Clones IST11858E10 (tigm) IST11589E8 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 1% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000679207 (Chr2:169458671..169459279 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAGTAGAGCGATGCCAAGGA Chr2:169459230..169459249 59.98 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000679207 (Chr2:169458671..169459279 +)
Downstram Exon
ENSMUSE00000638957 (Chr2:169616466..169616488 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAGTAGAGCGATGCCAAGGA Chr2:169459230..169459249 59.98 50 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000638958 Chr2:169458671..169459246 AGAAAGACGAGCGAGACTGC Chr2:169459160..169459179 59.9 55
upstream ENSMUSE00000679207 Chr2:169458671..169459279 AAGTAGAGCGATGCCAAGGA Chr2:169459230..169459249 59.98 50
upstream ENSMUSE00000679199 Chr2:169459176..169459279 AAGTAGAGCGATGCCAAGGA Chr2:169459230..169459249 59.98 50
upstream ENSMUSE00000679210 Chr2:169459240..169459279 No primer for this exon

*** Putative Vector Insertion (Chr 2: 169459280 - 169616465) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000638957 Chr2:169616466..169616488 No primer for this exon
downstream ENSMUSE00000638956 Chr2:169709013..169712086 GTGGAGACTTCGCTCGAAAC Chr2:169711486..169711505 60 55
downstream ENSMUSE00000549280 Chr2:169709026..169709460 CCCGACCAGTAGGAATCTGA Chr2:169709410..169709429 60.06 55
downstream ENSMUSE00000679198 Chr2:169709026..169714004 GTGGAGACTTCGCTCGAAAC Chr2:169711486..169711505 60 55
downstream ENSMUSE00000679205 Chr2:169709026..169712086 GTGGAGACTTCGCTCGAAAC Chr2:169711486..169711505 60 55
downstream ENSMUSE00000549278 Chr2:169709668..169712078 GTGGAGACTTCGCTCGAAAC Chr2:169711486..169711505 60 55
downstream ENSMUSE00000638955 Chr2:169738108..169738594 TGGCTTTCAGTCTCGTGTTG Chr2:169738144..169738163 60.03 50
downstream ENSMUSE00000679203 Chr2:169738108..169738593 TGGCTTTCAGTCTCGTGTTG Chr2:169738144..169738163 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AACGGAGGCAGGGAGTTAAT Chr2:169513315..169513335 59.96 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGCAAGTGTAGGGGAAAGC Chr2:169513312..169513332 59.74 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000047907