Part epidemic tissues of the fifth-instar larval mutants of four genes (white, brown, ok, lightoid) or the thorax and abdomen of adults (white, scarlet) and their corresponding wild types were dissected in phosphate buffer saline and then used to extract genomic DNA using TreliefTMAnimal Genomic DNA Kit (TsingKe, China) following the manufacturer’s protocols. The tissues of each individual were as a biological sample. At least three replicates were carried out for mutants of each gene. Except some adult mutants of white gene, part tissues of the same individual for the mutants and wild-types are also used for RNA extraction as described in the following part. Subsequently, primers flanking the target sites for each gene (Additional file 1: Table S4) were designed, and the PCR reaction were carried out using the 20 μl volumes, according to TransDirect PCR SuperMix (Trans, China). PCR products were TA-cloned into PMD19 vectors (Takara, Japan) and 10 clones were randomly picked up and sequenced for each individual. Sequence data were analyzed using SeqMan software (DNASTAR7.0) to determine the exact mutation type.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.