The lab tests and its results to rule out the presence of hepatitis A infection in the given patient. Concept introduction : To detect a known RNA sequence, methods such as reverse transcriptase polymerase chain reaction (RT-PCR) or RNA sequencing can be used. In both the cases, mRNA is first converted to complementary DNA (cDNA) and then analyzed. Hepatitis A is categorized as an RNA virus. Hepatitis A infection can be detected in the blood or tissue through these methods.

Question

1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results? 2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab? 3) Why are the cells incubated at 42°C?

Answer 1

Question

Chapter 20.4, Problem 3CC

Summary Introduction

To determine: The lab tests and its results to rule out the presence of hepatitis A infection in the given patient.

Concept introduction:

To detect a known RNA sequence, methods such as reverse transcriptase polymerase chain reaction (RT-PCR) or RNA sequencing can be used. In both the cases, mRNA is first converted to complementary DNA (cDNA) and then analyzed. Hepatitis A is categorized as an RNA virus. Hepatitis A infection can be detected in the blood or tissue through these methods.

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1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results? 2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab? 3) Why are the cells incubated at 42°C?

Overview of Transformation Protocol -Prepare competent bacteria for transformation: Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure. Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed. -Transformation procedure: Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA. Add CaCl2 to both tubes. Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube. Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes. Add recovery broth to both tubes. Incubate both tubes in a 37 C water bath for 5 minutes. Questions: 1) What differences would you expect to see between the…

Overview of Transformation Protocol -Prepare competent bacteria for transformation: Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure. Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed. -Transformation procedure: Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA. Add CaCl2 to both tubes. Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube. Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes. Add recovery broth to both tubes. Incubate both tubes in a 37 C water bath for 5 minutes. Questions: 1)What is the selectable marker in this experiment? How…