Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI27665
Trapped Gene
Wbp4 (ENSMUSG00000022023)
Vector Insertion
Chr 14: 79876912 - 79880179
Public Clones not available
Private Clones OST54161 (lexicon)
Severity of mutation (?) Insertion after 7% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000123036 (Chr14:79880180..79880252 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGCCGACTACTGGAAGTCAC Chr14:79880233..79880252 59.73 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000123036 (Chr14:79880180..79880252 -)
Downstram Exon
ENSMUSE00000123034 (Chr14:79876849..79876911 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGCCGACTACTGGAAGTCAC Chr14:79880233..79880252 59.73 60 ATCCTCCTGGCCACATTTTC Chr14:79876834..79876853 61.22 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000447775 Chr14:79880738..79881075 TATACCGACCCCACCTCTCA Chr14:79880939..79880958 60.33 55
upstream ENSMUSE00000123036 Chr14:79880180..79880252 GGCCGACTACTGGAAGTCAC Chr14:79880233..79880252 59.73 60

*** Putative Vector Insertion (Chr 14: 79876912 - 79880179) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000123034 Chr14:79876849..79876911 ATCCTCCTGGCCACATTTTC Chr14:79876834..79876853 61.22 50
downstream ENSMUSE00000123033 Chr14:79876610..79876739 ATCCTCTTGGTACGCCTTCA Chr14:79876619..79876638 59.69 50
downstream ENSMUSE00000123030 Chr14:79872159..79872332 GTGGGTTGGACAGTGCTGAT Chr14:79872262..79872281 61 55
downstream ENSMUSE00000123037 Chr14:79870494..79870540 CTGGCTTCTCCCATTGAGAT Chr14:79870498..79870517 59.24 50
downstream ENSMUSE00000123032 Chr14:79869912..79869987 No primer for this exon
downstream ENSMUSE00000123031 Chr14:79867080..79867270 AGGCCTCTCCAGCTTTCTTC Chr14:79867093..79867112 60.1 55
downstream ENSMUSE00000123029 Chr14:79865990..79866156 CTGGGCCAATTCTTTTCTCA Chr14:79866098..79866117 60.18 45
downstream ENSMUSE00000647202 Chr14:79859744..79861936 CTGGCAGACAAGAGGGTCTC Chr14:79860033..79860052 59.99 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr14:79880108..79880128 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACTTCGTGACTGGGAAAACC Chr14:79880112..79880132 59.04 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022023