Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6799
Trapped Gene
Zfp60 (ENSMUSG00000037640)
Vector Insertion
Chr 7: 28526075 - 28533186
Public Clones RHA043 (baygenomics) IST14703B10 (tigm) IST10854E6 (tigm) IST10750D4 (tigm)
Private Clones OST426710 (lexicon) OST359946 (lexicon)
Severity of mutation (?) Insertion after 12% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000597847 (Chr7:28525976..28526074 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAGGAGAAAGAGCCCTGGAT Chr7:28526020..28526039 60.72 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000597847 (Chr7:28525976..28526074 +)
Downstram Exon
ENSMUSE00000563275 (Chr7:28533187..28536708 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAGGAGAAAGAGCCCTGGAT Chr7:28526020..28526039 60.72 55 AGGCTGGAGACACGCTTAAA Chr7:28534501..28534520 60.01 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000676347 Chr7:28516408..28516489 TCGGGTCTTTCTGGTCTGAG Chr7:28516436..28516455 60.38 55
upstream ENSMUSE00000676350 Chr7:28516428..28516683 GTTTAGGGACGGTGCTGTGT Chr7:28516556..28516575 60.03 55
upstream ENSMUSE00000597846 Chr7:28521955..28522007 CCTGAAGAAGAATGGCCAAC Chr7:28521964..28521983 59.67 50
upstream ENSMUSE00000713205 Chr7:28521955..28522007 CCTGAAGAAGAATGGCCAAC Chr7:28521964..28521983 59.67 50
upstream ENSMUSE00000597848 Chr7:28523322..28523448 AGGGATGTGGCTGTTGACTT Chr7:28523337..28523356 59.58 50
upstream ENSMUSE00000597847 Chr7:28525976..28526074 CAGGAGAAAGAGCCCTGGAT Chr7:28526020..28526039 60.72 55

*** Putative Vector Insertion (Chr 7: 28526075 - 28533186) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000563275 Chr7:28533187..28536708 AGGCTGGAGACACGCTTAAA Chr7:28534501..28534520 60.01 50
downstream ENSMUSE00000676346 Chr7:28533187..28538721 AGGCTGGAGACACGCTTAAA Chr7:28534501..28534520 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTCAGGCACAGATGTGGTC Chr7:28526106..28526126 60.76 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAGATATTTCGTGACTGGGAAA Chr7:28529117..28529139 59.09 40.91 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000037640