Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31091
Trapped Gene
Tmlhe (ENSMUSG00000079834)
Vector Insertion
Chr NT_165789: 159774 - 216735
Public Clones (sanger) IST10099A11 (tigm) IST13583E7 (tigm) IST12527G10 (tigm) IST14529D4 (tigm)
IST14388F8 (tigm) IST13806H4 (tigm) IST11934B11 (tigm) IST13720E11 (tigm)
IST14173G5 (tigm) IST11095G7 (tigm) IST14588C1 (tigm) IST10985B8 (tigm)
IST11192D9 (tigm) IST14568A7 (tigm) IST14487E3 (tigm) IST13119H10 (tigm)
IST14515A9 (tigm) IST10099A11 (tigm) IST12400B12 (tigm) IST13539E7 (tigm)
IST14174A6 (tigm) IST14252G3 (tigm) IST13291A11 (tigm) IST14031F6 (tigm)
IST14828F2 (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
ENSMUSE00000704952 (ChrNT_165789:159647..159773 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGACAAAGCGGCACTACAG ChrNT_165789:159698..159717 59.66 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000704952 (ChrNT_165789:159647..159773 +)
Downstram Exon
ENSMUSE00000704951 (ChrNT_165789:216736..216917 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGACAAAGCGGCACTACAG ChrNT_165789:159698..159717 59.66 55 TCAAATTACGGAGCCTGCTT ChrNT_165789:216788..216807 59.85 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000704952 ChrNT_165789:159647..159773 CTGACAAAGCGGCACTACAG ChrNT_165789:159698..159717 59.66 55

*** Putative Vector Insertion (Chr NT_165789: 159774 - 216735) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000704951 ChrNT_165789:216736..216917 TCAAATTACGGAGCCTGCTT ChrNT_165789:216788..216807 59.85 45
downstream ENSMUSE00000704943 ChrNT_165789:258247..258423 GCTCCGCTGGTGAGTCTTAG ChrNT_165789:258350..258369 60.16 60
downstream ENSMUSE00000704940 ChrNT_165789:272714..272993 TCAGTGGGAGGGACATTTTC ChrNT_165789:272954..272973 59.9 50
downstream ENSMUSE00000704936 ChrNT_165789:285396..285515 ATGCAGTGTCACCTCTGGAA ChrNT_165789:285460..285479 59.26 50
downstream ENSMUSE00000704934 ChrNT_165789:326114..326350 ACCACCTGTTCCTTCATGCT ChrNT_165789:326159..326178 59.58 50
downstream ENSMUSE00000704928 ChrNT_165789:332450..332588 GTTAGGGTTCGGTGTGCTGT ChrNT_165789:332536..332555 60.03 55
downstream ENSMUSE00000704922 ChrNT_165789:333195..334143 CAGCCACAGAGTTGACGGTA ChrNT_165789:333271..333290 59.9 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTTTGTACCTAATCGCCTTGC ChrNT_165789:207816..207837 59.27 42.86 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TACCCGTGACTGGGAAAACC ChrNT_165789:207821..207841 62.06 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000079834