Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI30226
Trapped Gene
Pou2f1 (ENSMUSG00000026565)
Vector Insertion
Chr 1: 167864934 - 167932766
Public Clones (sanger) D072E01 (ggtc) D072E01 (ggtc) IST14276E10 (tigm) IST14225A7 (tigm)
IST13878E12 (tigm) IST14123F8 (tigm) IST13094B2 (tigm) IST14968H2 (tigm)
IST14278C4 (tigm) IST14188D1 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000711034 (Chr1:167932693..167932765 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGGAGCAGCGAGTCAAGATG Chr1:167932723..167932742 60.7 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000711034 (Chr1:167932693..167932765 -)
Downstram Exon
ENSMUSE00000492174 (Chr1:167864935..167865031 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGGAGCAGCGAGTCAAGATG Chr1:167932723..167932742 60.7 55 CAGTCACTGCAGTCCAGCAT Chr1:167864921..167864940 60.06 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000711034 Chr1:167932693..167932765 AGGAGCAGCGAGTCAAGATG Chr1:167932723..167932742 60.7 55
upstream ENSMUSE00000720487 Chr1:167932693..167932765 AGGAGCAGCGAGTCAAGATG Chr1:167932723..167932742 60.7 55
upstream ENSMUSE00000492174 Chr1:167864935..167865031 CGCTACCTGTTCCTTCTTGC Chr1:167864999..167865018 60.01 55

*** Putative Vector Insertion (Chr 1: 167864934 - 167932766) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000593540 Chr1:167861793..167861858 CTGTGCTGGCATCTTCACTC Chr1:167861771..167861790 59.58 55
downstream ENSMUSE00000593541 Chr1:167861793..167861858 CTGTGCTGGCATCTTCACTC Chr1:167861771..167861790 59.58 55
downstream ENSMUSE00000370820 Chr1:167847053..167847153 GGTGAAGATGTCCGAGGAAG Chr1:167847033..167847052 59.65 55
downstream ENSMUSE00000160566 Chr1:167845241..167845294 GACTGAGCAGCAGCCTGTAA Chr1:167845232..167845251 59.34 55
downstream ENSMUSE00000160578 Chr1:167843358..167843480 CGAATCTCCCGACTCTTCAC Chr1:167843429..167843448 59.8 55
downstream ENSMUSE00000160565 Chr1:167841419..167841607 TGCAATCTGTATGGGCTGAG Chr1:167841400..167841419 59.82 50
downstream ENSMUSE00000160582 Chr1:167839011..167839137 GCACCAACACAAACTGTTGC Chr1:167839052..167839071 60.21 50
downstream ENSMUSE00000379802 Chr1:167836816..167836910 AAAAGATTTTGCGCTTGCAG Chr1:167836864..167836883 60.51 40
downstream ENSMUSE00000226562 Chr1:167832271..167832444 TCAATTCGCTTTGGTGTTGA Chr1:167832343..167832362 60.23 40
downstream ENSMUSE00000536077 Chr1:167827756..167827897 GGTGGTTTGGCTGAAGTCAT Chr1:167827819..167827838 59.97 50
downstream ENSMUSE00000160572 Chr1:167826568..167826713 CTCTAAGGCCACACGGATGT Chr1:167826561..167826580 60.13 55
downstream ENSMUSE00000226538 Chr1:167825036..167825215 TCTTCCGAGGTAGGCTTTTG Chr1:167825171..167825190 59.45 50
downstream ENSMUSE00000226532 Chr1:167821941..167822046 AAAGGGAGGACAGGGTTGAC Chr1:167821954..167821973 60.35 55
downstream ENSMUSE00000226487 Chr1:167813178..167813249 TGCTGTGGTGGTGACTCTGT Chr1:167813157..167813176 60.37 55
downstream ENSMUSE00000500291 Chr1:167810261..167810606 GGAGAGGGACTCAAGGAAGG Chr1:167810485..167810504 60.19 60
downstream ENSMUSE00000358113 Chr1:167807108..167807196 TGCCAGTGTACTGTTGCTCA Chr1:167807096..167807115 59.03 50
downstream ENSMUSE00000687831 Chr1:167806034..167806107 ACAGAGGTGCAGTCACGATG Chr1:167806053..167806072 59.9 55
downstream ENSMUSE00000687828 Chr1:167805618..167805647 TCTCCAATCCATGAAGCAATC Chr1:167805605..167805625 60.03 42.86
downstream ENSMUSE00000335805 Chr1:167805616..167806179 ACAGAGGTGCAGTCACGATG Chr1:167806053..167806072 59.9 55
downstream ENSMUSE00000497207 Chr1:167805616..167806179 ACAGAGGTGCAGTCACGATG Chr1:167806053..167806072 59.9 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTTTCCTCCCCTTCCTCCTA Chr1:167911739..167911759 60 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTTTCCTCCCCTTCCTCCTA Chr1:167911739..167911759 60 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026565