Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI17223
Trapped Gene
Ppp1r2 (ENSMUSG00000047714)
Vector Insertion
Chr 16: 31265499 - 31274931
Public Clones (sanger) (sanger) P126C12 (ggtc) P126C12 (ggtc)
Private Clones OST447420 (lexicon) OST356682 (lexicon) OST356657 (lexicon) OST189227 (lexicon)
Severity of mutation (?) Insertion after 21% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000652940 (Chr16:31274932..31275078 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CGATCGTGGAAGAGGAACTG Chr16:31274934..31274953 60.79 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000652940 (Chr16:31274932..31275078 -)
Downstram Exon
ENSMUSE00000423486 (Chr16:31265391..31265498 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CGATCGTGGAAGAGGAACTG Chr16:31274934..31274953 60.79 55 GGGATGATATGTTGCCAGGA Chr16:31265425..31265444 60.69 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000466452 Chr16:31274932..31275363 CGATCGTGGAAGAGGAACTG Chr16:31274934..31274953 60.79 55
upstream ENSMUSE00000652940 Chr16:31274932..31275078 CGATCGTGGAAGAGGAACTG Chr16:31274934..31274953 60.79 55

*** Putative Vector Insertion (Chr 16: 31265499 - 31274931) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000423486 Chr16:31265391..31265498 GGGATGATATGTTGCCAGGA Chr16:31265425..31265444 60.69 50
downstream ENSMUSE00000423475 Chr16:31261316..31261393 TAGGCGTCTTCATCGTCACC Chr16:31261345..31261364 61.21 55
downstream ENSMUSE00000423470 Chr16:31260660..31260754 CTCCCGAGTCCGATACTTTG Chr16:31260687..31260706 59.69 55
downstream ENSMUSE00000423462 Chr16:31258423..31258590 TTCATGCTGTCTCCATCTGC Chr16:31258421..31258440 59.95 50
downstream ENSMUSE00000702118 Chr16:31258061..31258590 AGCACAGGAGGCACTGTCTT Chr16:31258335..31258354 60.06 55
downstream ENSMUSE00000339230 Chr16:31251627..31254819 ACCCACCTCCATGGTTACAA Chr16:31252200..31252219 60.09 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATTAATCGCCTTGCAGCACA Chr16:31265862..31265882 61.69 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGAGGTGAGTGAGGGAGAGC Chr16:31268914..31268934 60.55 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 ATAATCGCCTTGCAGCACAT Chr16:31266009..31266029 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CATCCCTTCTCCTCCGGAAC Chr16:31269100..31269120 63.18 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000047714