Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI37393
Trapped Gene
Btk (ENSMUSG00000031264)
Vector Insertion
Chr X: 131076908 - 131080179
Public Clones PST22185-NL (escells)
Private Clones not available
Severity of mutation (?) Insertion after 95% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000207747 (ChrX:131080021..131080178 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGGAAGATGCCGTATGAGAG ChrX:131080130..131080149 59.65 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000207747 (ChrX:131080021..131080178 -)
Downstram Exon
ENSMUSE00000694797 (ChrX:131076909..131077346 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGGAAGATGCCGTATGAGAG ChrX:131080130..131080149 59.65 55 GTATGCCAGGGAACGAGAGA ChrX:131077086..131077105 60.22 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000622602 ChrX:131117550..131117655 CTGGGGAGCTACCTGCATTA ChrX:131117563..131117582 60.23 55
upstream ENSMUSE00000694799 ChrX:131117550..131117667 CTGGGGAGCTACCTGCATTA ChrX:131117563..131117582 60.23 55
upstream ENSMUSE00000694798 ChrX:131112009..131112163 GGATTGCATCCTAAGGAAAGG ChrX:131112029..131112049 59.92 47.62
upstream ENSMUSE00000277353 ChrX:131108456..131108626 CTTCAAGAAGCGCCTGTTTC ChrX:131108506..131108525 60.13 50
upstream ENSMUSE00000712195 ChrX:131108456..131108626 CTTCAAGAAGCGCCTGTTTC ChrX:131108506..131108525 60.13 50
upstream ENSMUSE00000207746 ChrX:131107748..131107846 AAAAATCCCCCACCAGAAAG ChrX:131107758..131107777 60.16 45
upstream ENSMUSE00000207739 ChrX:131103444..131103512 AAAGGTTCCCGTACCCATTC ChrX:131103447..131103466 60.05 50
upstream ENSMUSE00000207741 ChrX:131100136..131100217 TTTCTCCCCAACTGAAGAGC ChrX:131100169..131100188 59.4 50
upstream ENSMUSE00000277330 ChrX:131093737..131093865 TGCTTCTGGATTGATGGACA ChrX:131093805..131093824 60.2 45
upstream ENSMUSE00000277322 ChrX:131093226..131093293 CCTGGGAGTTCTCATCGAAA ChrX:131093266..131093285 60.19 50
upstream ENSMUSE00000207752 ChrX:131091327..131091514 CAACCGAGCTGAAAAAGGTC ChrX:131091446..131091465 59.85 50
upstream ENSMUSE00000207745 ChrX:131090877..131090939 CAGGAAGGCTACATCCCAAGT ChrX:131090918..131090938 60.5 52.38
upstream ENSMUSE00000207740 ChrX:131090088..131090142 TCCAAGCACATGACTCGAAG ChrX:131090116..131090135 59.98 50
upstream ENSMUSE00000207736 ChrX:131089228..131089307 ACACCGTGTCTGTGTTTGCT ChrX:131089239..131089258 59.23 50
upstream ENSMUSE00000694803 ChrX:131089002..131089065 GTGATCCGCCATTACGTTGT ChrX:131089033..131089052 60.79 50
upstream ENSMUSE00000207748 ChrX:131088938..131089065 CGCCATTACGTTGTGTGTTC ChrX:131089027..131089046 60.03 50
upstream ENSMUSE00000207750 ChrX:131085741..131085815 AACAAAAACGCGCCTTCTAC ChrX:131085758..131085777 59.41 45
upstream ENSMUSE00000207751 ChrX:131084846..131085017 TTCCATGTCGGAGGATGAAT ChrX:131084871..131084890 60.28 45
upstream ENSMUSE00000207743 ChrX:131083954..131084170 TTGGAGTCGAAGCAGTTCCT ChrX:131083967..131083986 59.99 50
upstream ENSMUSE00000207753 ChrX:131082101..131082165 No primer for this exon
upstream ENSMUSE00000207742 ChrX:131081251..131081369 GGTGGTCTCCACCAGAAGTG ChrX:131081297..131081316 60.56 60
upstream ENSMUSE00000207747 ChrX:131080021..131080178 GGGAAGATGCCGTATGAGAG ChrX:131080130..131080149 59.65 55
upstream ENSMUSE00000694802 ChrX:131076980..131077346 TCTCTCGTTCCCTGGCATAC ChrX:131077108..131077127 60.22 55
upstream ENSMUSE00000368789 ChrX:131076914..131077346 TCTCTCGTTCCCTGGCATAC ChrX:131077108..131077127 60.22 55
upstream ENSMUSE00000694797 ChrX:131076909..131077346 TCTCTCGTTCCCTGGCATAC ChrX:131077108..131077127 60.22 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAAGGATCCCAAGAAAGCTG ChrX:131080204..131080224 59.81 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAAGGATCCCAAGAAAGCTG ChrX:131080204..131080224 59.81 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031264