Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI28977
Trapped Gene
Mgrn1 (ENSMUSG00000022517)
Vector Insertion
Chr 16: 4927902 - 4930690
Public Clones not available
Private Clones OST14450 (lexicon)
Severity of mutation (?) Insertion after 81% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000127768 (Chr16:4927675..4927901 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTATGAGCCCATCTCCTTGC Chr16:4927698..4927717 59.8 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000127768 (Chr16:4927675..4927901 +)
Downstram Exon
ENSMUSE00000127767 (Chr16:4930691..4930814 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTATGAGCCCATCTCCTTGC Chr16:4927698..4927717 59.8 55 TGAGAGCTTCTCCTCGTCCT Chr16:4930748..4930767 59.28 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000622259 Chr16:4886317..4886473 GGCGTAGAGGACATCGACAT Chr16:4886416..4886435 60.1 55
upstream ENSMUSE00000622258 Chr16:4907365..4907486 ATTTGACACTCCCCATCCTG Chr16:4907408..4907427 59.78 50
upstream ENSMUSE00000622256 Chr16:4907368..4907486 ATTTGACACTCCCCATCCTG Chr16:4907408..4907427 59.78 50
upstream ENSMUSE00000127772 Chr16:4910720..4910808 AGACTCTCTCCGGCTTGTGA Chr16:4910788..4910807 60.13 55
upstream ENSMUSE00000127776 Chr16:4911322..4911468 ATTGTCAGGCGGTAGAGGAG Chr16:4911429..4911448 59.31 55
upstream ENSMUSE00000127775 Chr16:4913690..4913807 TGACTTCTCCGAGTGGAAGG Chr16:4913780..4913799 60.38 55
upstream ENSMUSE00000127766 Chr16:4915753..4915819 GGGGTGTGTTTCCAGTAGTCA Chr16:4915772..4915792 59.88 52.38
upstream ENSMUSE00000127777 Chr16:4916105..4916154 No primer for this exon
upstream ENSMUSE00000127765 Chr16:4918866..4918913 CTCCGTGAAGCCATTAAAGC Chr16:4918883..4918902 59.85 50
upstream ENSMUSE00000127764 Chr16:4920313..4920381 ACGGCATCGAGAACAAGAAC Chr16:4920347..4920366 60.26 50
upstream ENSMUSE00000127773 Chr16:4922594..4922753 TCTGCACTTCCTGTGCTGAC Chr16:4922691..4922710 60.19 55
upstream ENSMUSE00000430591 Chr16:4924423..4924532 TCTCCTTCAGCCCTGTCTTG Chr16:4924483..4924502 60.52 55
upstream ENSMUSE00000430573 Chr16:4926309..4926374 TGGCCAGCAAGAAACCTAAA Chr16:4926346..4926365 60.74 45
upstream ENSMUSE00000127768 Chr16:4927675..4927901 CTATGAGCCCATCTCCTTGC Chr16:4927698..4927717 59.8 55

*** Putative Vector Insertion (Chr 16: 4927902 - 4930690) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000127767 Chr16:4930691..4930814 TGAGAGCTTCTCCTCGTCCT Chr16:4930748..4930767 59.28 55
downstream ENSMUSE00000127771 Chr16:4931113..4931155 GCTTTAGGGACGGCTCATCT Chr16:4931155..4931174 60.73 55
downstream ENSMUSE00000127779 Chr16:4932200..4932298 No primer for this exon
downstream ENSMUSE00000127778 Chr16:4934195..4934239 CAACAGAGCACGACTCAGGA Chr16:4934224..4934243 60.18 55
downstream ENSMUSE00000564234 Chr16:4936896..4938292 CAGGGCAGTTCCTTAAGCTG Chr16:4937941..4937960 60.01 55
downstream ENSMUSE00000622257 Chr16:4936896..4938292 CAGGGCAGTTCCTTAAGCTG Chr16:4937941..4937960 60.01 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGAAGGGCAAGACACAGAGC Chr16:4927869..4927889 59.6 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGAAGGGCAAGACACAGAGC Chr16:4927869..4927889 59.6 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022517