Answer
Step 1: Obtain a RNA sample
Step 2: Add one primer complementary to the 3' end of the RNA of interest. Also add the RT enzyme (reverse trasnscriptase) , dNTPs, and a buffer with Mg2+. Incubate the reaction appropriately.
Step 3: Degrade the RNA with base leaving just the cDNA
Step 4: cDNA is produced which is single stranded and complementary to the RNA of interest.
Step 5: Double stranded DNA for the region of interest is produced.
Step 6: Add two primers (forward and reverse) located within the sequence of the RNA of interest. Also add DNA polymerase, dNTPs, and a buffer with Mg2+. cycle temperature appropriately.
Question 10 0.5 pts Order the steps in RT-PCR: • Add one primer complementary to the...
Order the steps in RT-PCR: 1. Add one primer complementary to the 3' end of the RNA of interest. Also add the RT enzyme (reverse transcriptase), dNTPs, and a buffer with Mg2+. Incubate the reaction appropriately. 2. Degrade the RNA strand with base, leaving just the cDNA. 3. cDNA is produced, which is single-stranded and complementary to the RNA of interest. 4. Double-stranded DNA for the region of interest is produced. 5. Add two primers (one forward and one reverse)...
Question 11 0.5 pts Compare and contrast PCR techniques. Uses DNA polymerase Uses reverse transcriptase ✓ [ Choose] PCR and qPCR RT-PCR RT-PCR and qRT-PCR qPCR qRT-PCR None of the above PCR, RT-PCR, qPCR, and qRT-PCR qPCR and qRT-PCR Precisely quantitates nucleic acids in "real- time" Synthesizes RNA [Choose]
Carolina Savirana Craz 3/12/20 GECC-Polymerase Chain Reaction 1. What is the purpose of the polymerase chain reaction? a. To repair damaged DNA b. To make copies of entire chromosomes c. To make copies of specific regions of DNA d. To prepare cells for cell division 2. The polymerase chain reaction is most comparable to what cellular process? a. Mitosis b. Replication c. Transcription d. Translation 3. When enzymes are elongating (building) a newly synthesized DNA strand in PCR, new nucleotides...
QUESTION 1: You are inserting a gene into an MCS found within the LacZ gene. Using blue/white colony selection, why could you assume that white colonies have modified plasmids? a. A blue colony means the LacZ reading-frame was disrupted b. A blue colony means your gene has mutations c. A white colony means the LacZ reading-frame is intact d. A white colony means the LacZ reading-frame was disrupted QUESTION 2: You are performing a PCR using primers with a sequence perfectly...
1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteinase K; guanidine HCI; SDS 2. Why is the ratio of the OD at 260 and 280 nm used to estimate DNA purity? 3. In one paragraph, summarize basic principles of PCR technique in your own words. List all the reagents you will need to perform a PCR experiment. Does this method tell you what genetic modifications were made? If yes, describe how you can...