Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24423
Trapped Gene
Pnp1 (ENSMUSG00000021871)
Vector Insertion
Chr 14: 51570394 - 51570506
Public Clones not available
Private Clones OST221417 (lexicon)
Severity of mutation (?) Insertion after 53% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000617135 (Chr14:51570218..51570393 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGACTCAACCCCAATTTTGA Chr14:51570287..51570306 59.77 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000617135 (Chr14:51570218..51570393 +)
Downstram Exon
ENSMUSE00000617134 (Chr14:51570507..51570697 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGACTCAACCCCAATTTTGA Chr14:51570287..51570306 59.77 45 ATCCCGGTCATAAGCATCAG Chr14:51570555..51570574 59.92 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000289946 Chr14:51564212..51564268 No primer for this exon
upstream ENSMUSE00000688658 Chr14:51567258..51567265 No primer for this exon
upstream ENSMUSE00000560158 Chr14:51567527..51567696 GGCTTCTGCAACACACTGAA Chr14:51567562..51567581 60.03 50
upstream ENSMUSE00000617136 Chr14:51569933..51570036 CTGGTGTTTGGATTGCTGAA Chr14:51569953..51569972 59.69 45
upstream ENSMUSE00000617135 Chr14:51570218..51570393 GGACTCAACCCCAATTTTGA Chr14:51570287..51570306 59.77 45

*** Putative Vector Insertion (Chr 14: 51570394 - 51570506) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000617134 Chr14:51570507..51570697 ATCCCGGTCATAAGCATCAG Chr14:51570555..51570574 59.92 50
downstream ENSMUSE00000496127 Chr14:51571079..51571688 AGTGCCTTGCGACGATAACT Chr14:51571120..51571139 59.9 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTCTGGCCTTGTCCTAATCG Chr14:51570431..51570451 59.69 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGCAGAGAGTCTGGCCTTGT Chr14:51570423..51570443 61.15 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000021871