Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31821
Trapped Gene
l7Rn6 (ENSMUSG00000062797)
Vector Insertion
Chr 7: 97072073 - 97084475
Public Clones IST14731C5 (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
ENSMUSE00000477314 (Chr7:97084237..97084474 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGCATGGGAGGATCTGTCTA Chr7:97084341..97084360 60.03 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000477314 (Chr7:97084237..97084474 -)
Downstram Exon
ENSMUSE00000487322 (Chr7:97072074..97072225 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGCATGGGAGGATCTGTCTA Chr7:97084341..97084360 60.03 55 AAAGGAGTCAACGGAGGACA Chr7:97072058..97072077 59.7 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000489415 Chr7:97089479..97089695 TAGTGGAGGGTCACGTTCCT Chr7:97089619..97089638 59.57 55
upstream ENSMUSE00000672472 Chr7:97089479..97089594 No primer for this exon
upstream ENSMUSE00000713514 Chr7:97089479..97089709 GAGGCGTGCAAGTTAGGAAG Chr7:97089685..97089704 60.01 55
upstream ENSMUSE00000672471 Chr7:97088490..97088553 GGTTCCAAGAAACCATGGAC Chr7:97088524..97088543 59.24 50
upstream ENSMUSE00000477314 Chr7:97084237..97084474 GGCATGGGAGGATCTGTCTA Chr7:97084341..97084360 60.03 55
upstream ENSMUSE00000487322 Chr7:97072074..97072225 CCTCCGTTGACTCCTTTACG Chr7:97072077..97072096 59.73 55
upstream ENSMUSE00000713158 Chr7:97072074..97072225 CCTCCGTTGACTCCTTTACG Chr7:97072077..97072096 59.73 55

*** Putative Vector Insertion (Chr 7: 97072073 - 97084475) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000490457 Chr7:97068583..97068701 TGCTGGGATGAACATTTCTG Chr7:97068578..97068597 59.65 45
downstream ENSMUSE00000711507 Chr7:97067434..97067517 TTTCCAAAAGAGGGGGTTCT Chr7:97067447..97067466 59.91 45
downstream ENSMUSE00000672470 Chr7:97067430..97067517 TTTCCAAAAGAGGGGGTTCT Chr7:97067447..97067466 59.91 45
downstream ENSMUSE00000515882 Chr7:97067199..97067517 TTTCCAAAAGAGGGGGTTCT Chr7:97067447..97067466 59.91 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGGATTTTTAAATGGCAGTTCC Chr7:97072481..97072503 58.57 36.36 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGGATTTTTAAATGGCAGTTCC Chr7:97072481..97072503 58.57 36.36 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000062797