Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI8317
Trapped Gene
4632419K20Rik (ENSMUSG00000044465)
Vector Insertion
Chr 7: 112533937 - 112536697
Public Clones RRF380 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 41% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000289431 (Chr7:112536698..112536865 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTTCCTGCGATTCCTGTTGT Chr7:112536763..112536782 60.26 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000289431 (Chr7:112536698..112536865 -)
Downstram Exon
ENSMUSE00000289425 (Chr7:112533857..112533936 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTTCCTGCGATTCCTGTTGT Chr7:112536763..112536782 60.26 50 TGAGGTTCAGAAGGGTCCTG Chr7:112533869..112533888 60.23 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000436926 Chr7:112548398..112548525 CGTGGTGCAAGTGAAAGAGA Chr7:112548430..112548449 60.03 50
upstream ENSMUSE00000716976 Chr7:112548398..112548546 CGTGGTGCAAGTGAAAGAGA Chr7:112548430..112548449 60.03 50
upstream ENSMUSE00000719956 Chr7:112548398..112548522 CGTGGTGCAAGTGAAAGAGA Chr7:112548430..112548449 60.03 50
upstream ENSMUSE00000712218 Chr7:112540091..112540181 GCTGAGCCACATTCTGTGAA Chr7:112540134..112540153 59.99 50
upstream ENSMUSE00000436924 Chr7:112538535..112538866 AGACCGGATGAAGCAGATTG Chr7:112538754..112538773 60.22 50
upstream ENSMUSE00000716956 Chr7:112538535..112538866 AGACCGGATGAAGCAGATTG Chr7:112538754..112538773 60.22 50
upstream ENSMUSE00000503564 Chr7:112537768..112538406 AGGCCTGGTGCTACTTCTCA Chr7:112538004..112538023 60.01 55
upstream ENSMUSE00000289443 Chr7:112537293..112537451 TGCCTCGAAAGATTGAGGTT Chr7:112537395..112537414 59.81 45
upstream ENSMUSE00000289435 Chr7:112537070..112537156 TTCACAATGGTTTCCTGGTG Chr7:112537097..112537116 59.39 45
upstream ENSMUSE00000289431 Chr7:112536698..112536865 CTTCCTGCGATTCCTGTTGT Chr7:112536763..112536782 60.26 50

*** Putative Vector Insertion (Chr 7: 112533937 - 112536697) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000289425 Chr7:112533857..112533936 TGAGGTTCAGAAGGGTCCTG Chr7:112533869..112533888 60.23 55
downstream ENSMUSE00000289417 Chr7:112533502..112533665 AAAGATCCACATCACGCACA Chr7:112533579..112533598 60.12 45
downstream ENSMUSE00000716496 Chr7:112533460..112533665 AAAGATCCACATCACGCACA Chr7:112533579..112533598 60.12 45
downstream ENSMUSE00000720002 Chr7:112532889..112533086 GAGCCACCTAGGCTCTGTTG Chr7:112532959..112532978 60.01 60
downstream ENSMUSE00000289404 Chr7:112532295..112533086 GAGCCACCTAGGCTCTGTTG Chr7:112532959..112532978 60.01 60
downstream ENSMUSE00000671188 Chr7:112532290..112533086 GAGCCACCTAGGCTCTGTTG Chr7:112532959..112532978 60.01 60
downstream ENSMUSE00000289393 Chr7:112529913..112530091 TGTTGGTGTTGAGCAGGAAA Chr7:112529926..112529945 60.28 45
downstream ENSMUSE00000436436 Chr7:112529594..112529756 TGGATAGCAGAGCAGGGAAG Chr7:112529662..112529681 60.49 55
downstream ENSMUSE00000506337 Chr7:112529594..112529732 TGGATAGCAGAGCAGGGAAG Chr7:112529662..112529681 60.49 55
downstream ENSMUSE00000715253 Chr7:112527126..112527643 AGTAATTCCCCCAAGGATGG Chr7:112527588..112527607 60.01 50
downstream ENSMUSE00000384683 Chr7:112527125..112527643 AGTAATTCCCCCAAGGATGG Chr7:112527588..112527607 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCTGGGTGAGCCATCTTTTA Chr7:112536644..112536664 60.07 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGAGCCATCTTTCGTGACTG Chr7:112536638..112536658 59.98 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000044465