Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI34742
Trapped Gene
Rp2h (ENSMUSG00000060090)
Vector Insertion
Chr X: 19958946 - 19974346
Public Clones IST15086F6 (tigm) IST13968F4 (tigm) IST13968F4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 82% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000396064 (ChrX:19958831..19958945 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAAAACTGAAGACGCCCAAA ChrX:19958878..19958897 60.23 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000396064 (ChrX:19958831..19958945 +)
Downstram Exon
ENSMUSE00000358246 (ChrX:19974347..19974432 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAAAACTGAAGACGCCCAAA ChrX:19958878..19958897 60.23 45 AATTCCAAGGCAATCACAGG ChrX:19974371..19974390 59.93 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000702688 ChrX:19941607..19941954 ATTGGTTGCCATAGGTCGTC ChrX:19941654..19941673 59.82 50
upstream ENSMUSE00000702680 ChrX:19941685..19941954 AAAGGTGTCTGAGCGAGTGG ChrX:19941812..19941831 60.44 55
upstream ENSMUSE00000441005 ChrX:19941692..19941954 AAAGGTGTCTGAGCGAGTGG ChrX:19941812..19941831 60.44 55
upstream ENSMUSE00000557122 ChrX:19941701..19941954 AAAGGTGTCTGAGCGAGTGG ChrX:19941812..19941831 60.44 55
upstream ENSMUSE00000702677 ChrX:19942166..19942324 GGAAGGTGCTGAATCTCCAA ChrX:19942220..19942239 60.19 50
upstream ENSMUSE00000205775 ChrX:19954007..19954672 AGAAAATGCCGTGGTTCAAG ChrX:19954474..19954493 60.11 45
upstream ENSMUSE00000396064 ChrX:19958831..19958945 GAAAACTGAAGACGCCCAAA ChrX:19958878..19958897 60.23 45

*** Putative Vector Insertion (Chr X: 19958946 - 19974346) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000358246 ChrX:19974347..19974432 AATTCCAAGGCAATCACAGG ChrX:19974371..19974390 59.93 45
downstream ENSMUSE00000702684 ChrX:19974521..19974529 No primer for this exon
downstream ENSMUSE00000582156 ChrX:19974778..19977979 GTTGGCCTGAAAGTTTCCAA ChrX:19976451..19976470 60.09 45
downstream ENSMUSE00000624677 ChrX:19974778..19977254 ACATCTCCAGATGCGGTCTC ChrX:19974821..19974840 60.23 55
downstream ENSMUSE00000702687 ChrX:19974778..19974872 ACATCTCCAGATGCGGTCTC ChrX:19974821..19974840 60.23 55
downstream ENSMUSE00000702686 ChrX:19977896..19977983 TGCTTGTTCTCAATGCTCCA ChrX:19977943..19977962 60.54 45
downstream ENSMUSE00000440987 ChrX:19981175..19981205 CTAGCAAGGTCCTGGGTTCC ChrX:19981208..19981227 61.01 60
downstream ENSMUSE00000440981 ChrX:19981302..19981370 AGGCCATCCAAACTTTCTGA ChrX:19981340..19981359 59.67 45
downstream ENSMUSE00000656601 ChrX:19982334..19982450 AGTTCAGGGGATCCAATGCT ChrX:19982366..19982385 60.85 50
downstream ENSMUSE00000702689 ChrX:19982573..19982781 TTCCAGCCTTGCTCAAAAGT ChrX:19982691..19982710 59.99 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTTCATTAATCGCCTTGCAG ChrX:19961991..19962011 58.39 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGTTCATCGTGACTGGGAAA ChrX:19961990..19962010 60.09 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000060090