Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI7061
Trapped Gene
8430426H19Rik (ENSMUSG00000055228)
Vector Insertion
Chr 13: 62558108 - 62558292
Public Clones RRZ059 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000576353 (Chr13:62558293..62558419 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGAGGACGTGCATGTGAACT Chr13:62558386..62558405 60.32 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000576353 (Chr13:62558293..62558419 -)
Downstram Exon
ENSMUSE00000576351 (Chr13:62558047..62558107 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGAGGACGTGCATGTGAACT Chr13:62558386..62558405 60.32 50 ATGGGCTTCCCAATTAAAGC Chr13:62558066..62558085 60.28 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000681633 Chr13:62568067..62568149 AATTGCCAGCGATCTACGAC Chr13:62568078..62568097 60.24 50
upstream ENSMUSE00000576353 Chr13:62558293..62558419 TGAGGACGTGCATGTGAACT Chr13:62558386..62558405 60.32 50

*** Putative Vector Insertion (Chr 13: 62558108 - 62558292) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000576351 Chr13:62558047..62558107 ATGGGCTTCCCAATTAAAGC Chr13:62558066..62558085 60.28 45
downstream ENSMUSE00000484388 Chr13:62556336..62556553 GAATCCTTTCATGCCTTTGG Chr13:62556428..62556447 59.51 45
downstream ENSMUSE00000576349 Chr13:62555683..62556333 ATGACCGTATTGGGAAAAGG Chr13:62555694..62555713 58.76 45
downstream ENSMUSE00000521021 Chr13:62554834..62556553 GGATTCTGGCCCCTACAAAT Chr13:62554939..62554958 60.15 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr13:62558221..62558241 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector

Come back to gene ENSMUSG00000055228