Problem Set due May 4, 2021 23:00 HKT Past due

Buffers
You are a sophomore at MIT in course 7 (biology). In addition to taking 7.05 Biochemistry in the
spring semester, you have enrolled in a biochemistry project lab, an intensive laboratory subject (a
class at MITC that focuses on conducting various experiments to master the basic biochemical
techniques. You take the subject to prepare to conduct biochemical research at an MIT lab during
the summer (as part of the UROP program).

You walk into the project lab and sit down at a lab bench. A few minutes later, your friend Dorothy,
also a course 7 sophomore, joins you at the bench. She too is planning on participating in the
UROP program over the summer. You chat with each other as other students file into the lab. You
recognize a few faces from class with Professor Yaffe.

After waiting the customary five minutes past class start time, the teaching assistants (TAs) Eva
and Daly introduce themselves and explain that today’s class will focus on making buffers. You
and your classmates audibly groan. Eva, chuckling, acknowledges that the first few labs will be a
bit boring, but that she and Daly will try to keep things interesting by making connections to
current research and applications. Daly reminds you and your classmates that buffers seem
modest, but are quite important, and using the incorrect ones can be disastrous for experiments
in the lab, or in real life.

General Approach to Making Buffers

0.0/1.0 point (graded)
Eva asks if you remember the general approach for making a good buffer mentioned in class. You
raise your hand to answer.

To make a good buffer, you need to mix a Select an option acid with its
Select an option in roughly equal molar amounts. Ideally, the should be
Select an option the value of the weak acid.

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Alternative Approaches to Making Buffers

0.0/1.0 point (graded)
Cesar, a classmate from your 7.05 recitation (a small review session held by a TAC, raises his hand.
He asks if there are other approaches to make buffers. Daly nods and asks you all what you think.
Which of the following methods would also result in a good buffer? Select all that apply.

Starting with a strong acid in solution, add weak base until the is equal to the .

Starting with a weak acid in solution, add strong base until the is equal to the .

Starting with a strong base in solution, add weak acid until the is equal to the .

Starting with a weak base in solution, add strong acid until the is equal to the .

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Use of Buffers in Tissue Culture

0.0/1.0 point (graded)
Daly wants to elaborate on the importance of buffers in lab and in life before the students start
making anything. As a 4th year graduate student who utilizes tissue culture techniques to study
cancer biology, she uses growth media - liquid containing nutrients and buffer that allow isolated
cells to survive and grow. She shows you and your peers commercial and lab-made growth media
and buffers commonly used in tissue culture.

A typical buffer incorporated in cell growth media is HEPES (a Good buffer) at . Cells are
incubated at !, at which the of HEPES is . What is the ratio of acid to conjugate base
in such a buffer? Enter your answer as a number (with two significant figures) in the space
provided.
Hint: pay close attention to your ratio!

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pH Indicators in Tissue Culture

0.0/1.0 point (graded)
You notice and ask why some of the commercial cell growth media are visibly red. Daly explains
this is because the media contain a indicator, a dye that changes color over a specific
range.

Suppose you want a pH indicator in your growth medium containing HEPES buffer to alert you to
when the pH begins to drop below physiological due to cells releasing metabolites (small
molecules resulting from the breakdown of nutrients available in the growth medium). Which of
these indicators would you include in your growth medium? Select the best answer.
these indicators would you include in your growth medium? Select the best answer.

an indicator dye that changes color at

an indicator dye that changes color at

an indicator dye that changes color at

an indicator dye that changes color at

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Topic: Problem Set: Buffers and Amino Acids / Buffers

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! Certificate and past due

2
I missed due dates :( Can I still earn a certificate ?