This material may consist of step-by-step explanations on how to solve a problem or examples of proper writing, including the use of citations, references, bibliographies, and formatting. This material is made available for the sole purpose of studying and learning - misuse is strictly forbidden.1. In an organic extraction, what would happen if you forgot to put SDS into the lysis buffer?
Answer: Sodium dodecyl sulfate (SDS) is an anionic detergent that helps in chemical lysis of cell membranes. It totally denatures protein including DNAse and RNAse enzymes as well as membrane proteins. SDS helps in emulsification of membrane lipids and disrupts the polar interactions with proteins. It forms complexes with proteins and lipids that cause precipitation of these materials out of the solution. So, if SDS is not added into lysis buffer, yield of extracted DNA will be low and quality will be poor too due to presence of nucleases.
2. With the FTA punch-and-wash method, the DNA remains on the punch, even during subsequent reactions, such as amplification. Do you think that one punch (washed) can be used for both a real— time PCR quantitation & human identification locus amplification? Why or why not?
Answer: The same punch cannot be used for both real-time PCR quantitation & human identification locus amplification.
After real-time PCR, huge amount of amplified dsDNA will remain attached on the punch that will interfere the binding of probe during human identification locus amplification....