Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33946
Trapped Gene
Plekhc1 (ENSMUSG00000037712)
Vector Insertion
Chr 14: 46088914 - 46092555
Public Clones IST14716H4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 47% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000314872 (Chr14:46092447..46092554 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGAGATCGAATGCACGGAAG Chr14:46092476..46092495 60.36 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000314872 (Chr14:46092447..46092554 -)
Downstram Exon
ENSMUSE00000314864 (Chr14:46088915..46089049 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGAGATCGAATGCACGGAAG Chr14:46092476..46092495 60.36 50 GCAGCGTCAACTTCATCAAC Chr14:46088945..46088964 59.45 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000453533 Chr14:46149611..46149713 No primer for this exon
upstream ENSMUSE00000706602 Chr14:46149609..46149793 No primer for this exon
upstream ENSMUSE00000408319 Chr14:46149100..46149266 GCTCTGGACGGGATAAGGAT Chr14:46149234..46149253 60.43 55
upstream ENSMUSE00000722177 Chr14:46149100..46149266 GCTCTGGACGGGATAAGGAT Chr14:46149234..46149253 60.43 55
upstream ENSMUSE00000315015 Chr14:46124172..46124405 TGGATAAGTGTGGCATCCAG Chr14:46124308..46124327 59.52 50
upstream ENSMUSE00000314890 Chr14:46102107..46102241 CCCAGAGATCCCACAAAGAA Chr14:46102184..46102203 60.04 50
upstream ENSMUSE00000314881 Chr14:46095563..46095788 ACCAACTTCTGCCTGGTTTG Chr14:46095690..46095709 60.15 50
upstream ENSMUSE00000315007 Chr14:46095248..46095350 TTGCTGCTCCGATTCAAGTA Chr14:46095276..46095295 59.57 45
upstream ENSMUSE00000314872 Chr14:46092447..46092554 AGAGATCGAATGCACGGAAG Chr14:46092476..46092495 60.36 50
upstream ENSMUSE00000314864 Chr14:46088915..46089049 GCCCTTTCTGACCTGGAGAT Chr14:46088949..46088968 60.6 55

*** Putative Vector Insertion (Chr 14: 46088914 - 46092555) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000314991 Chr14:46088178..46088227 TCAGCAAGTTCTGGGATTGA Chr14:46088174..46088193 59.37 45
downstream ENSMUSE00000314985 Chr14:46087013..46087137 AGCAGGAGATGGACGTGTCT Chr14:46087045..46087064 59.87 55
downstream ENSMUSE00000314978 Chr14:46084759..46084865 ACAGCGAAGCCAGATCTCAT Chr14:46084743..46084762 59.98 50
downstream ENSMUSE00000314969 Chr14:46084399..46084620 TCAGATGCTGCATTTTCAGG Chr14:46084475..46084494 59.95 45
downstream ENSMUSE00000314959 Chr14:46081791..46081915 GCCAGGCTTGAATGAATCTC Chr14:46081809..46081828 59.78 50
downstream ENSMUSE00000314950 Chr14:46081442..46081583 GTCCATCCGAATCAACCTGT Chr14:46081498..46081517 59.79 50
downstream ENSMUSE00000706596 Chr14:46079428..46079698 ATTTTGGTCTTTCGCACGAG Chr14:46079554..46079573 60.25 45
downstream ENSMUSE00000336558 Chr14:46078467..46079698 ATTTTGGTCTTTCGCACGAG Chr14:46079554..46079573 60.25 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTGTTACAGGCCGTGTCTCC Chr14:46092565..46092585 60.58 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGTTACAGGCCGTGTCTCC Chr14:46092565..46092585 60.58 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000037712