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ZKO number ZKO986
Recent progress of this line F2 individuals have been indentified
Gene name mdh1a
Ensembl ID ENSDARG00000017772
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGTGCTGGATGGAGTCGTCAtgg
Cas9 target site 2 sequence
Position in the gene exon 4
Restriction enzyme (RE) Hin fI
PCR primer (forward) tggagtgtactgctcggctaa
PCR primer (reverse) aaccaaccagggaagaatgat
Length of expected PCR product (bp) 311
Length of expected RE digestion product (bp) 167+144
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO 986
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 40
Survival rate of F0 embryo @ 1dpf (%) 90.6
Method to estimate mutation efficiency Restriction enzyme digestionHin fI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO986.pdf
Mutation efficiency in F0 embryos (%) 30
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 20%(1/5)
Wild-type (WT) sequence TGCTGCCGGTGCTGGATGGAGTCGTCATGGAGCTGCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TGCTGCCGGTGCTGGATGGAG-----------CTGCA
F0-2 TGCTGCCGGTGCTGGATGGATGGAGTCGTTTCATGGAGCTGCA
Allele name and predicted product 986a
986b
TGCTGCCGGTGCTGGATGGAG-----------CTGCA; Truncated protein(67aa)
TGCTGCCGGTGCTGGATGGATGGAGTCGTTTCATGGAGCTGCA; Extended protein(355aa)
突变创建时间 2014-07-02
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information:
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