Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI12165
Trapped Gene
Nme1 (ENSMUSG00000037601)
Vector Insertion
Chr 11: 93829534 - 93829575
Public Clones W227C01 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000376631 (Chr11:93829535..93829574 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCGGTAAAGCCTTGTCATCT Chr11:93829541..93829560 59.34 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000376631 (Chr11:93829535..93829574 -)
Downstram Exon
ENSMUSE00000674586 (Chr11:93829535..93829586 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCGGTAAAGCCTTGTCATCT Chr11:93829541..93829560 59.34 50 TCAGATGACAAGGCTTTACCG Chr11:93829517..93829537 60.26 47.62

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000376631 Chr11:93829535..93829574 GCGGTAAAGCCTTGTCATCT Chr11:93829541..93829560 59.34 50
upstream ENSMUSE00000674586 Chr11:93829535..93829586 GCGGTAAAGCCTTGTCATCT Chr11:93829541..93829560 59.34 50

*** Putative Vector Insertion (Chr 11: 93829534 - 93829575) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000110585 Chr11:93827122..93827251 CTCGAACCGCTTGATGATCT Chr11:93827139..93827158 60.36 50
downstream ENSMUSE00000713683 Chr11:93827122..93827251 CTCGAACCGCTTGATGATCT Chr11:93827139..93827158 60.36 50
downstream ENSMUSE00000110582 Chr11:93824541..93824642 AGCAACCACTGGTCCTGAGT Chr11:93824522..93824541 59.76 55
downstream ENSMUSE00000674585 Chr11:93822023..93822149 GGTTGGTCTCTCCAAGCATC Chr11:93822015..93822034 59.66 55
downstream ENSMUSE00000110586 Chr11:93821982..93822094 CTCGTATGGTCCCAGGCTTA Chr11:93821985..93822004 60.09 55
downstream ENSMUSE00000382394 Chr11:93820547..93820827 AGTGGCACGTAACACTGCAC Chr11:93820549..93820568 59.83 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTGAAGGGGTAATCGCCTTG Chr11:93829521..93829541 61.86 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTCATCTGAAGGGGCGTGAC Chr11:93829526..93829546 63.02 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000037601