Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI10820
Trapped Gene
Bclaf1 (ENSMUSG00000037608)
Vector Insertion
Chr 10: 20048014 - 20049115
Public Clones P114E07 (ggtc)
Private Clones OST214681 (lexicon)
Severity of mutation (?) Insertion after 87% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000615776 (Chr10:20047908..20048013 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACCTGCAGTGACCCTAAACG Chr10:20047924..20047943 60.17 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000615776 (Chr10:20047908..20048013 +)
Downstram Exon
ENSMUSE00000615775 (Chr10:20049116..20049200 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACCTGCAGTGACCCTAAACG Chr10:20047924..20047943 60.17 55 CGCTGGGAGAGATGTCAATC Chr10:20049141..20049160 60.77 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000484704 Chr10:20032275..20032410 GAAATGGCCGCTGTGTAGTT Chr10:20032345..20032364 60.14 50
upstream ENSMUSE00000705467 Chr10:20032278..20032410 GAAATGGCCGCTGTGTAGTT Chr10:20032345..20032364 60.14 50
upstream ENSMUSE00000099625 Chr10:20037814..20037917 No primer for this exon
upstream ENSMUSE00000099624 Chr10:20041809..20041922 GCCGCTCCAATTCTAGATCA Chr10:20041823..20041842 60.32 50
upstream ENSMUSE00000412631 Chr10:20042769..20043674 TCACATCCCATCACGAAGAA Chr10:20043540..20043559 60.05 45
upstream ENSMUSE00000487284 Chr10:20042775..20043674 TCACATCCCATCACGAAGAA Chr10:20043540..20043559 60.05 45
upstream ENSMUSE00000615778 Chr10:20044946..20045611 CCAGTGATTCTCACCGTCCT Chr10:20045545..20045564 60.11 55
upstream ENSMUSE00000615777 Chr10:20045736..20045905 TAGCACGCTTGTCCATTCTG Chr10:20045769..20045788 60.01 50
upstream ENSMUSE00000615776 Chr10:20047908..20048013 ACCTGCAGTGACCCTAAACG Chr10:20047924..20047943 60.17 55

*** Putative Vector Insertion (Chr 10: 20048014 - 20049115) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000615775 Chr10:20049116..20049200 CGCTGGGAGAGATGTCAATC Chr10:20049141..20049160 60.77 55
downstream ENSMUSE00000615774 Chr10:20051892..20052067 GGAATCTCCCCGCTCTTTAC Chr10:20051984..20052003 60.04 55
downstream ENSMUSE00000577177 Chr10:20052202..20052363 TCCAACCTCCTGATGTAGCC Chr10:20052333..20052352 60.07 55
downstream ENSMUSE00000577184 Chr10:20053198..20053378 ATGGTCCCGCTCTCTACCTT Chr10:20053228..20053247 60.1 55
downstream ENSMUSE00000705466 Chr10:20053198..20053378 ATGGTCCCGCTCTCTACCTT Chr10:20053228..20053247 60.1 55
downstream ENSMUSE00000577183 Chr10:20054305..20054451 TGTCCCAGCAAAAACTCCTC Chr10:20054355..20054374 60.23 50
downstream ENSMUSE00000577180 Chr10:20059515..20059727 TTCCTCTTTTGGCCCAGTAA Chr10:20059563..20059582 59.68 45
downstream ENSMUSE00000705465 Chr10:20059515..20059727 TTCCTCTTTTGGCCCAGTAA Chr10:20059563..20059582 59.68 45
downstream ENSMUSE00000577179 Chr10:20059858..20062307 AGGGGAGCATCATGCAATAC Chr10:20061332..20061351 59.92 50
downstream ENSMUSE00000705464 Chr10:20059858..20060072 TCTGCTCCCTGTTGCAATCT Chr10:20059891..20059910 60.94 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCCCCGAGATACACAGGTA Chr10:20047998..20048018 59.57 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ATGCTCGTGACTGGGAAAAC Chr10:20048060..20048080 60.12 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000037608