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ZKO number ZKO844
Recent progress of this line F2 individuals have been indentified
Gene name POLD4
Ensembl ID ENSDARG00000016994(1)
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGCTGAAGAACTTTGATC
Cas9 target site 2 sequence
Position in the gene Exon3
Restriction enzyme (RE) MboI
PCR primer (forward) TTCAGATATGACACCAAAGCGT
PCR primer (reverse) AGAAATGACAACAGGCCAAGC
Length of expected PCR product (bp) 464
Length of expected RE digestion product (bp) 307bp+157bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO 844
Dosage of injected Cas9 mRNA (ng/ul) 250
Dosage of injected gRNA (ng/ul) 175
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency Restriction enzyme digestionMboI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO844.pdf
Mutation efficiency in F0 embryos (%) 85
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 70.1
Wild-type (WT) sequence AAAGAGAAAAAGATATACAGGAGCTGAAGAACTTTGATCTGGATTGGAAGTTTGGACCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 AAAGAGAAAAAGATATACAGGAGCTGAAGAACTTTG - - - - -GATTGGAAGTTTGGACCA
F0-2 AAAGAGAAAAAGATATACAGGAGCTGAAGAACTTTGGATTGGAACAGGTAA- - - - - - - - -
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 AAAGAGAAAAAGATATACAGGAGCTGAAGAACTTTG - - - - -GATTGGAAGTTTGGACCA
F0-2 AAAGAGAAAAAGATATACAGGAGCTGAAGAACTTTGGATTGGAACAGGTAA- - - - - - - - -
Allele name and predicted product 844a
844b
(-5,+0)
(-26,+15)
突变创建时间 2014-03-19
填表时间 2013/12/19
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information:
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