Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33765
Trapped Gene
Plekhc1 (ENSMUSG00000037712)
Vector Insertion
Chr 14: 46084758 - 46087138
Public Clones (sanger) (sanger) IST11497H2 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 63% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000314985 (Chr14:46087013..46087137 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGACACGTCCATCTCCTGCT Chr14:46087067..46087086 59.87 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000314985 (Chr14:46087013..46087137 -)
Downstram Exon
ENSMUSE00000314978 (Chr14:46084759..46084865 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGACACGTCCATCTCCTGCT Chr14:46087067..46087086 59.87 55 ACAGCGAAGCCAGATCTCAT Chr14:46084743..46084762 59.98 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
upstream ENSMUSE00000314991 Chr14:46088178..46088227 TCAATCCCAGAACTTGCTGA Chr14:46088196..46088215 59.37 45
upstream ENSMUSE00000314985 Chr14:46087013..46087137 AGACACGTCCATCTCCTGCT Chr14:46087067..46087086 59.87 55
upstream ENSMUSE00000314978 Chr14:46084759..46084865 ATGAGATCTGGCTTCGCTGT Chr14:46084765..46084784 59.98 50

*** Putative Vector Insertion (Chr 14: 46084758 - 46087138) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
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 CTTACAGGCCGAAGAAGCTG Chr14:46087123..46087143 60.15 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTTACAGGCCGAAGAAGCTG Chr14:46087123..46087143 60.15 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000037712