Complete the following tasks. You are fresh recruits to a molecular biology laboratory, and your new boss has tasked you to study mutations in NRAS, a gene that she suspects is involved in cancer pathogenesis. Task A: Polymerase Chain Reaction Master Mix Your first task is to isolate and amplify the NRAS gene from cDNA extracted from different samples through PCR. You will need to run a total of 7 PCRs: 3 normal samples, 3 cancerous samples, and 1 negative control. To make things easier in the lab, when running, multiple reactions, the components are prepared not individually, but as a master mix-all the components for multiple reactions are prepared in bulk, except for the template DNA, which is added separately once the master mix has been distributed into individual tubes. The table below lists the different components for PCR, the available stock concentrations of these components, and the needed working concentrations for the PCR itself. Complete the table by supplying the needed volumes of each component for a single reaction and for the master mix (the first row has been filled in as an example). PCR Component Buffer for Taq polymerase Forward primer Reverse primer dNTPs Taq polymerase Sterile distilled deionized water Template Stock concentration 10X 40 uM 10 uM 25 mM 200 U/ul n/a Total 400 ng/ul Total Working concentration 1X 1.0 uM 1.0 um 1 mM 1 U/ul (to fill reaction volume to 10 ul) 40 ng/ul Volume for 1 reaction (µl.) 1.0 Volume for 7 reactions (μl) 10.0 7.0 9.0 ul 63.0 ul 1.0 (added separately to each reaction after master mix is distributed to different tubes) 70.0

Biol370 - Assignment #2 – Spring 2020: Replication - Transcription - Translation Due 3/10/2020 (Before the beginning of the class) Name ID Instruction: 1. Print this document (US-letter only) Fill the table below with the appropriate color (see coloring rule) 3. Complete and annotate the drawing in attachment (see page #2). Make sure all the listed terms are present in your final drawing! (graded using 5 randomly selected terms 2. This type of drawing makes only sense for Eukaryotes/Prokaryotes (circle one) because: or The annotation must follow the coloring rule : DNA in BLACK. RNA in RED. Proteins and AAs in GREEN. Complex structures with some DNA and/or RNA and/or Proteins in BLUE. Color Color Color DNA DNA pol III Stop codon RNA RNA polymerase 5'-UTR Peptide Helicase 3'-UTR Leading strand Topoisomerase CDS Lagging strand Ligase RBS Coding strand Primer Ribosome SSU Template strand -10/-35 Box Ribosome LSU 5'/3' ends (all) Transcription start AA-TRNA N-/C- terminal ends…

Question:- How many different types of alterations in chromosomal numbers are there? -How can I use the F plasmid to introduce the transfer of the URA+ gene from a wild type strain into the ura- mutant strain you isolated in Q5.   -After obtaining the ura- mutant, I want to transform this ura- mutant back to wild type with a DNA sample prepared from the wild type cell. How can I achieve this transformation experimentally?

Problem 4:  You are planning an experiment to use zinc finger nucleases to generate a double-stranded DNA break in a targeted sequence of DNA Name the protein domain in a zinc finger nuclease that is responsible for generating the DNA break Fok1 Name two other genome-engineering proteins that could be used for generating the double-stranded DNA break   What are the advantages and disadvantages of each strategy?