Bio 1A Lab 3

Front 1

What is the purpose of using the chemical DCPIP

Back 1

DCPIP (blue color) is equivalence to NADP+ (clear color)

**we use DCPIP to see photosynthesis happens b/c as it get reduces to DCPIPH2 it goes from blue to clear. Whereas NADP+ is clear and NADPH+ is also clear

Front 2

When DCPIP get reduces down to DCPIPH2 it becomes clear? what does clear indicates

Back 2

the clear your solution becomes the more sugar you make

Front 3

How do you mess up photosynthesis in this experiment?

Back 3

(1) by adding DCMU or
(2) add Methylamine

Front 4

what does DCMU do? and what are the consequences

Back 4

DCMU blocks ETC
**stops transfer of e- from PS II to PS I
**shut off bingin H+
**so it will slowly decrease photosynthesis because we won't be able to make NADPH+

Front 5

what does methylamine do? and what are the consequences

Back 5

Methylamine will go into the lumen and protonate all the H+ (suck u all the H+ in lumen)
**so cell speeds up ETC to pump more H+ however not making any ATP to make sugar.

____ + NADPH + H+ ------> CH2O + NADP+ (can't make it w/o atp)

**can't make carbon fixation anymore; need energy (ATP) for rxn to happen.

**in this case we won't be able to make anymore NADP+ either

Front 6

[pigment extraction and chromatography]

1. solvent is (hydrophobic or hydrophillic)

2. chromotography paper is (hydrophilic or hydrophobic)

Back 6

**solvent is hydrophobic

**chrom. paper is hydrophillic

Front 7

Polar pigments will parttion more with the ____ and migrate ___(a shorter or longer distance)

Back 7

hydrophillic paper, migrate a shorter distance

Front 8

hydrophobic/nonpolar pigments will parttion more with the ____ and migrate ___(a shorter or longer distance)

Back 8

developing solvent and migrate further.

Front 9

List from order of low Rf values to large Rf values

*Chlorophyll a, chlorophyll b, and cartanoid

Back 9

Chloro. a, b, and cartanoid

Front 10

what is the hill reaction?

Back 10

Hill rxn involves the light-dependent transfer of e- from H2O to an acceptor that results in the evolution of O2.

**substitute DCPIP as artificial e- acceptor
**Rate photosynthesis by the rate of color change

Front 11

What is use in the calvin cycle/carbon fixation

Back 11

ATP + CO2+ NADPH+ + H+ -------> CH2O + NADP+

Front 12

describes how Methylamine (CH3NH2) affect these:

1. ATP production
2. NADPH production
3. CO2 fixation

Back 12

1) ATP production decrease
2) NADPH production increase
3) CO2 fixation decrease (need both ATP and NADH)

Front 13

describes how Dark control affect these pH of stroma (or buffer) is at pH=8.2, and the pH of the thylakoid lumen is buffered at pH=4:

1. ATP production
2. NADPH production
3. CO2 fixation

Back 13

1.) ATP production-- no change. There is a H+ gradient to make ATP
2.) NADPH decrease b/c no light to excite an e-
3.) CO2 fixation--- decrease

Front 14

describes how DCMU + CH3NH2 affect these:

1. ATP production
2. NADPH production
3. CO2 fixation

Back 14

1. Decrease
2.decrease
3. CO2 fixation decrease

Front 15

What is the purpose of dark control?

Back 15

to check to make sure that DCPIP isn't reduced by reagents in your solution. If it is, then you need to subtract this reduction from your experimental results to get the actual results due to e- transport.

Front 16

Trace the path of e-'s through the Z scheme (the non-cyclic light)

Back 16

e- from water ----> PS II primary e- acceptor, which is considered the first member of the 1st ETC by some people ---> PQ to Cyt b_6f complex (H+ pump) ---> PC to PSC chl a -----> to PS I primary e- Acceptor, which is considered to be the first member of the 2nd ETC by some people to Fd (in the middle of 2nd ETC) to NADP+ reductase (at the end of the 2nd ETC) ---> to NADP+ to NADPH

Front 17

O----

*what does the symbol/oreintatiib above mean
**and where is it located

Back 17

**It represents ATP synthase
**The O is on the stroma side of the thylakoid membrane---> this represents the globular portion of the ATP synthase enzyme and is in the thylakoid membrane. --- this represent the proton channel.

Front 18

where is light first absorbed in a photosystem??

Back 18

antenna complex/antenna pigment molecules in both photosystems

Front 19

How does osmoticum prevent lysis?

Back 19

Here, the osmoticum is sucrose. Enough sucrose is dissolved in P buffer to make it isotonic to the stroma. the sucrose P buffer is isotonic to the stroma "solution"-- the chloroplasts are resuspended in a solution that is isotonic to it, meaning it has the same # of particles per unit volume as the stroma. So there is no NET water moving in or not (the # of water molecules diffuse into stroma= # of particles per unit volume as the stroma.) so the ct's remain intact (not shrink/crenate and not swell to point where they're lysed.0

Front 20

1 um (micrometer) = ____ meters

1 mm= ____ meters

Back 20

1X10^-6

0.001 or 1X10^-3

Front 21

For our Hill rxn: Light controls (is our positive (+) control)

what is the purpose of the light control or what are you checking?

Back 21

**(+) control we expect to see something.

**we are checking our chloroplast prep, to see a "normal" rate of DCPIP reduction.

Front 22

What if you see nothing in your in the light control?

Back 22

**something is worng, maybe with your chloroplast prep. Manybe the reagents your using are not working, or you forgot to add them, etc.

Front 23

Dark control is our negative control (where we expect to see nothing)

what are we checking? and what if we see something?

Back 23

dark control we expect to see nothing. thus were

**we are checking to see any "baseline" reduction of DCPIP

**If we see something-- meaning DCPIP either reduced itself automatically or there is something causing DCPIP reduction automatically.