Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31447
Trapped Gene
Rgs7 (ENSMUSG00000026527)
Vector Insertion
Chr 1: 177023934 - 177051941
Public Clones IST11108A2 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000434759 (Chr1:177051859..177051940 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGAGCATTCTGGGATGTCCA Chr1:177051869..177051888 60.62 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000434759 (Chr1:177051859..177051940 -)
Downstram Exon
ENSMUSE00000323486 (Chr1:177023935..177024009 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGAGCATTCTGGGATGTCCA Chr1:177051869..177051888 60.62 50 TTCGTGTTTTGTGTGGGTTT Chr1:177023915..177023934 58.92 40

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000500975 Chr1:177421184..177421261 TTATGGACAGACCAGCAACG Chr1:177421225..177421244 59.72 50
upstream ENSMUSE00000498969 Chr1:177200948..177201044 GCACGTATGCAAGACGAGAA Chr1:177201010..177201029 60.02 50
upstream ENSMUSE00000686997 Chr1:177200948..177201046 GCACGTATGCAAGACGAGAA Chr1:177201010..177201029 60.02 50
upstream ENSMUSE00000505410 Chr1:177119308..177119358 No primer for this exon
upstream ENSMUSE00000434784 Chr1:177083255..177083361 GCACCTTCTACCGGTTTCAA Chr1:177083255..177083274 60.11 50
upstream ENSMUSE00000686991 Chr1:177083255..177083362 GCACCTTCTACCGGTTTCAA Chr1:177083255..177083274 60.11 50
upstream ENSMUSE00000434776 Chr1:177079783..177079834 TTTTTGGCCGTCAAATTGTT Chr1:177079807..177079826 60.33 35
upstream ENSMUSE00000434768 Chr1:177053062..177053126 ACAATGCAAAACAAGGCAAG Chr1:177053087..177053106 58.82 40
upstream ENSMUSE00000535194 Chr1:177052026..177052102 CCGGAAGTGGGAGTTCATTT Chr1:177052051..177052070 61.24 50
upstream ENSMUSE00000434759 Chr1:177051859..177051940 AGAGCATTCTGGGATGTCCA Chr1:177051869..177051888 60.62 50
upstream ENSMUSE00000323486 Chr1:177023935..177024009 AACCCACACAAAACACGAAA Chr1:177023936..177023955 58.92 40

*** Putative Vector Insertion (Chr 1: 177023934 - 177051941) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000160233 Chr1:177021125..177021223 GGCTTGGTTTCTGGTGTAGG Chr1:177021131..177021150 59.59 55
downstream ENSMUSE00000160232 Chr1:177019550..177019611 No primer for this exon
downstream ENSMUSE00000160228 Chr1:177016247..177016357 GGCACAAGAAACGGGTCATA Chr1:177016285..177016304 60.89 50
downstream ENSMUSE00000658629 Chr1:177016247..177016311 CTGAGAGCCATGGATTGGAA Chr1:177016256..177016275 61.15 50
downstream ENSMUSE00000160231 Chr1:177014103..177014228 AAAACCCCATCGTTTTACCC Chr1:177014167..177014186 59.92 45
downstream ENSMUSE00000160225 Chr1:177009536..177009722 TCGGATAGGCCTTCTCTTCA Chr1:177009658..177009677 59.91 50
downstream ENSMUSE00000535186 Chr1:177008320..177008409 GGGGGTATGAGTCGCTCTTC Chr1:177008348..177008367 60.99 60
downstream ENSMUSE00000658628 Chr1:177006955..177007008 AGAGACGTTCTGCGATCCAT Chr1:177006955..177006974 59.83 50
downstream ENSMUSE00000658624 Chr1:177006148..177006228 GCATGGGAAAATAGGGAGGT Chr1:177006180..177006199 60.15 50
downstream ENSMUSE00000535184 Chr1:176996772..176996825 No primer for this exon
downstream ENSMUSE00000658627 Chr1:176996748..176996825 TTGAGGACAGGAGCTTGCTT Chr1:176996731..176996750 60.13 50
downstream ENSMUSE00000535185 Chr1:176989875..176989925 No primer for this exon
downstream ENSMUSE00000658626 Chr1:176989867..176989925 No primer for this exon
downstream ENSMUSE00000686995 Chr1:176989348..176989925 CAAGGGTGAAACGACACCTT Chr1:176989395..176989414 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCTGCCTCTGAACCACATTG Chr1:177030937..177030957 59.83 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GATCACTCAGGTCCACATTTCA Chr1:177030919..177030941 59.97 45.46 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026527