Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI10748
Trapped Gene
Ltbp4 (ENSMUSG00000040488)
Vector Insertion
Chr 7: 28094087 - 28094304
Public Clones G029B04 (ggtc) CMHD-GT_354G10-3 (cmhd) IST11001C7 (tigm) IST13899C6 (tigm)
IST14882F4 (tigm) IST14543H3 (tigm) IST10997C11 (tigm) IST15061G4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 82% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000535645 (Chr7:28094305..28094433 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGGACGAATGCCAACTCTT Chr7:28094412..28094431 60.12 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000535645 (Chr7:28094305..28094433 -)
Downstram Exon
ENSMUSE00000535644 (Chr7:28093940..28094086 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGGACGAATGCCAACTCTT Chr7:28094412..28094431 60.12 50 GCTCGCAGGTACAGTGGTAAG Chr7:28093978..28093998 59.95 57.14

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000501397 Chr7:28122447..28122711 AGAAAGTGAGTCGGCCTCTG Chr7:28122681..28122700 59.6 55
upstream ENSMUSE00000676489 Chr7:28120696..28120730 TGTCACTGCTGCCTAGACCA Chr7:28120701..28120720 60.62 55
upstream ENSMUSE00000488113 Chr7:28120672..28120730 GCTGCCTAGACCAGACACCT Chr7:28120694..28120713 59.48 60
upstream ENSMUSE00000490043 Chr7:28120416..28120554 CAGTCCCGTGGAGAAGAGTC Chr7:28120444..28120463 59.83 60
upstream ENSMUSE00000484703 Chr7:28119856..28119965 GAAAAGAAGGCCCCCACATC Chr7:28119880..28119899 62.97 55
upstream ENSMUSE00000676487 Chr7:28118431..28118648 CTCTGGGTGTCGCTATTGGT Chr7:28118583..28118602 60.13 55
upstream ENSMUSE00000676486 Chr7:28118374..28118428 No primer for this exon
upstream ENSMUSE00000708828 Chr7:28118374..28118667 CTCTGGGTGTCGCTATTGGT Chr7:28118583..28118602 60.13 55
upstream ENSMUSE00000709573 Chr7:28118374..28118632 CTCTGGGTGTCGCTATTGGT Chr7:28118583..28118602 60.13 55
upstream ENSMUSE00000716944 Chr7:28118374..28118682 CTCTGGGTGTCGCTATTGGT Chr7:28118583..28118602 60.13 55
upstream ENSMUSE00000307052 Chr7:28115146..28115337 TCCCTTGATCTGTCACAACG Chr7:28115311..28115330 59.68 50
upstream ENSMUSE00000535658 Chr7:28114590..28114837 TTTAGAGAACTGCGCGGAAG Chr7:28114594..28114613 60.65 50
upstream ENSMUSE00000535657 Chr7:28114398..28114500 CCTGGGGTGTTCATGATTGT Chr7:28114420..28114439 60.63 50
upstream ENSMUSE00000535656 Chr7:28113987..28114061 ACCCGGCTTCGAGAGAGTTA Chr7:28114002..28114021 61.28 55
upstream ENSMUSE00000535654 Chr7:28113759..28113881 GTGTATGTCCGGACGGTTTC Chr7:28113790..28113809 60.24 55
upstream ENSMUSE00000535653 Chr7:28113205..28113369 GCCAGCTCTGTCCACCTTAT Chr7:28113212..28113231 59.31 55
upstream ENSMUSE00000407351 Chr7:28112657..28112806 TGACCTCCGATACAACACCA Chr7:28112735..28112754 59.96 50
upstream ENSMUSE00000362610 Chr7:28112280..28112531 TTCTGCCTACTCGTCGACCT Chr7:28112509..28112528 60.01 55
upstream ENSMUSE00000307165 Chr7:28111711..28111842 CAGCATGTGTCAGCGAAATC Chr7:28111813..28111832 60.42 50
upstream ENSMUSE00000307108 Chr7:28111506..28111631 AATACACCAGGCAGCTTTCG Chr7:28111559..28111578 60.27 50
upstream ENSMUSE00000307161 Chr7:28110697..28110822 GCCAGGCAGTTTCCTATGTG Chr7:28110745..28110764 60.66 55
upstream ENSMUSE00000307156 Chr7:28110038..28110280 GCGTGTGAAGAAGATGTGGA Chr7:28110157..28110176 59.84 50
upstream ENSMUSE00000307950 Chr7:28109313..28109438 TTCAAGTGTGTCTGCCCAAC Chr7:28109351..28109370 59.73 50
upstream ENSMUSE00000308043 Chr7:28109106..28109225 GGGCAGGAGTGTGTGAACTC Chr7:28109171..28109190 60.72 60
upstream ENSMUSE00000308034 Chr7:28108161..28108289 CATGGCCAGTGCACAAATAC Chr7:28108232..28108251 59.99 50
upstream ENSMUSE00000308025 Chr7:28107830..28107955 ACCTCGTGGAGCATCTTGTC Chr7:28107833..28107852 60.27 55
upstream ENSMUSE00000308018 Chr7:28107308..28107430 AGAGCGGCATCTGTACCAAC Chr7:28107378..28107397 60.28 55
upstream ENSMUSE00000308012 Chr7:28107041..28107172 ACTGCGATCCAGGTTACCAC Chr7:28107066..28107085 60 55
upstream ENSMUSE00000307031 Chr7:28104644..28104775 AATACGGCTCTGCGATTTGT Chr7:28104738..28104757 59.74 45
upstream ENSMUSE00000307152 Chr7:28104430..28104555 CTTTCAGTGCGTCTGTGACC Chr7:28104469..28104488 59.47 55
upstream ENSMUSE00000535649 Chr7:28099256..28099402 CATGACTGGACGCTGTGTTC Chr7:28099274..28099293 60.32 55
upstream ENSMUSE00000711483 Chr7:28096579..28096831 GACCGGTTTCAATCCAGAAA Chr7:28096779..28096798 59.91 45
upstream ENSMUSE00000535648 Chr7:28095540..28095806 GACAATATCTTGGCCCGAAA Chr7:28095626..28095645 59.9 45
upstream ENSMUSE00000535647 Chr7:28095200..28095271 ACCAGTCATTGTGTCCTCACG Chr7:28095245..28095265 61.03 52.38
upstream ENSMUSE00000535645 Chr7:28094305..28094433 GTGGACGAATGCCAACTCTT Chr7:28094412..28094431 60.12 50

*** Putative Vector Insertion (Chr 7: 28094087 - 28094304) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000535644 Chr7:28093940..28094086 GCTCGCAGGTACAGTGGTAAG Chr7:28093978..28093998 59.95 57.14
downstream ENSMUSE00000307972 Chr7:28092787..28092939 AGACAGCAGCACTCGGTGTA Chr7:28092818..28092837 59.65 55
downstream ENSMUSE00000307021 Chr7:28091581..28091961 AGGGGTCGTAGGGTAGCACT Chr7:28091772..28091791 60.02 60
downstream ENSMUSE00000307074 Chr7:28091086..28091238 TACCACACTCTTCGGCCTCT Chr7:28091169..28091188 59.87 55
downstream ENSMUSE00000719934 Chr7:28091017..28091238 TACCACACTCTTCGGCCTCT Chr7:28091169..28091188 59.87 55
downstream ENSMUSE00000465029 Chr7:28090161..28090569 GCAAATCCTGGACGACAGAT Chr7:28090442..28090461 60.08 50
downstream ENSMUSE00000709638 Chr7:28090159..28090569 GCAAATCCTGGACGACAGAT Chr7:28090442..28090461 60.08 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCACCTTTTCCTGCAGATGT Chr7:28094428..28094448 60.11 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCACCTTTTCCTGCAGATGT Chr7:28094428..28094448 60.11 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000040488