Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32753
Trapped Gene
Manba (ENSMUSG00000028164)
Vector Insertion
Chr 3: 135174045 - 135174766
Public Clones IST13388G5 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 14% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000242249 (Chr3:135173939..135174044 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GACACGGTTGCAGAAATCCT Chr3:135173976..135173995 60.12 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000242249 (Chr3:135173939..135174044 +)
Downstram Exon
ENSMUSE00000242240 (Chr3:135174767..135174937 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GACACGGTTGCAGAAATCCT Chr3:135173976..135173995 60.12 50 GTGTGAGCCTTGCTCTGACA Chr3:135174867..135174886 60.19 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000242269 Chr3:135148578..135148821 AGCATGCATCTTCACCTGCT Chr3:135148642..135148661 60.98 50
upstream ENSMUSE00000242259 Chr3:135169873..135169967 GGACCTACAGCACGGAATTT Chr3:135169928..135169947 59.06 50
upstream ENSMUSE00000242249 Chr3:135173939..135174044 GACACGGTTGCAGAAATCCT Chr3:135173976..135173995 60.12 50

*** Putative Vector Insertion (Chr 3: 135174045 - 135174766) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000242240 Chr3:135174767..135174937 GTGTGAGCCTTGCTCTGACA Chr3:135174867..135174886 60.19 55
downstream ENSMUSE00000242234 Chr3:135180867..135180990 AGATTCCCTGAGAGGGGAAG Chr3:135180924..135180943 59.63 55
downstream ENSMUSE00000242226 Chr3:135185956..135186131 TTGAGGGATGGCTACTGTCA Chr3:135186056..135186075 59.24 50
downstream ENSMUSE00000176937 Chr3:135187481..135187591 CCTCCATCCAGAGCAAAGAG Chr3:135187569..135187588 59.94 55
downstream ENSMUSE00000176926 Chr3:135205276..135205427 AGTCGGCTGGAATCCAATTT Chr3:135205402..135205421 60.83 45
downstream ENSMUSE00000176931 Chr3:135207690..135207807 GAGTGTTCATGTTCGCATCC Chr3:135207739..135207758 59.09 50
downstream ENSMUSE00000446365 Chr3:135210514..135210600 TTTCCTCACGGAGGCTAAGA Chr3:135210588..135210607 59.95 50
downstream ENSMUSE00000176940 Chr3:135212164..135212331 ATGTCCCTGGGATTTACGTG Chr3:135212270..135212289 59.67 50
downstream ENSMUSE00000500705 Chr3:135214027..135214245 TGAAAGGACGGCTCTTGTCT Chr3:135214051..135214070 59.99 50
downstream ENSMUSE00000176942 Chr3:135217724..135217888 GGAGAAGCGGCTGTTGTAAG Chr3:135217768..135217787 60.01 55
downstream ENSMUSE00000414334 Chr3:135226169..135226313 GGCCTTTTCCGTCCACTATC Chr3:135226250..135226269 60.83 55
downstream ENSMUSE00000445781 Chr3:135229452..135229594 AACAGTGGAGCGAAGAAACG Chr3:135229515..135229534 60.43 50
downstream ENSMUSE00000242177 Chr3:135230428..135230685 CTCCGGCCTTAACCACAATA Chr3:135230506..135230525 59.95 50
downstream ENSMUSE00000414715 Chr3:135233179..135234293 AGTCTTCTGGCCGTGCTTTA Chr3:135233641..135233660 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCTGGCCTTTATGTTCGTG Chr3:135174073..135174093 60.27 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGCTGGCCTTTATGTTCGTG Chr3:135174073..135174093 60.27 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028164