Skip to main content

Lab Note #4 Second experiment

 Lab Note #4 Second experiment 

Since the previous result was not clear and cells did not get reduced as much as I thought, I decided to the second experiment by putting an increased concentration of PLA2 and 5FU on colorectal cells. 

Method:

1. I took plates with cells(WIDR and HCT15) out of a culture medium.

2. I got rid of liquid media using a suction machine. 

3. I put 500ul of TE buffer so that cells on the bottom of the plates float. After I put them on the centrifuge I waited for 5 minutes. (If the cells are on TE buffer too long, cells can die.) 

4. After 5 minutes, I slightly shook the plate with TE buffer and cells so that the cells are detached from the bottom of the plates. 

5. I put them on two 1.5ml tubes.

6. I put the tubes on a centrifuge that spin in a speed of 7500 rpm for 2 minutes. Then I can saw cells on the bottom of the tube separated from TE buffer. 

7. I sucked TE buffer carefully so that the cells don't get sucked up together. 

8. I put 200ul of RPMI and did a pipetting so that cells are mixed with RPMI evenly

9. I put RPMI + cells on 15ml tube (I made the liquid flow on the wall)

10. With a big pipette, I put 7 ml of RPMI and shook it up and down slightly.

11. I put 500ul of solution on a 24 wells plate. 

12. On the same 24 wells plate, I put PLA2 and 5FU according to the following concentration. 


 

 

1

2

3

4

5

6

 

PLA2

1500u/ml

0

5

10

20

40

50

u/ml

 

 

0

1.65

3.3

6.6

13.2

16.5

Ul

5FU

10mg/ml

0

200

400

600

800

 

uM

 

 

0

1.3

2.6

3.9

5.2

6.5

ul


After 72 hours, I checked the result using the same cell counting machine. 

 

HCT15

 

 

 

PLA2

0

5

10

20

Live Cell

2.20E+06

8.70E+05

1.07E+06

1.37E+06

Viability

93.3

93.2

94.8

96.9



 

HCT15

 

 

 

5FU

0

200

400

600

Live Cell

2.12E+06

2.02E+06

1.65E+06

1.49E+06

Viability

95.5

96.3

95.7

92

 

 

WIDR

 

 

 

PLA2

0

5

10

20

Live Cell

2.61E+06

3.55E+06

2.67E+06

2.44E+06

Viability

98.8

97.4

95.8

96.1

 

 

WIDR

 

 

 

5FU

0

200

400

600

Live Cell

2.15E+06

2.75E+05

5.70E+05

5.70E+05

Viability

97.6

80.6

86

86

Interpretation:

Half Life:

-22.3709

1709.09

10.60117

-824.385

I got minus half-lives, which proves that I could not get the result I wanted. 

So I decided to do the another experiment. 

Labels

Labels:

Comments

Post a Comment

Popular posts from this blog

Actually, it is a privilege!

             "There is always something we can learn from tradition" This is what my parents always say (that I cannot totally agree) whenever I complain about conservative, patriarchal tradition of Korean holiday, Chuseok and New years day. Every time I go to grandfather's house in Korean holiday, I feel disgust to see scene that shows radical patriarchal tradition of Korea. Only women work in kitchen as soon as they arrive in grandfather's house, and men watch TV in living room, seems not to even 'consider' helping women. However, women are the last to eat meal since they are allowed to eat after they finish cleaning a kitchen and has to set the men's table first. The difficulty that women go through when they eat does not end here. Since men use much larger table for eating even though they are fewer than women, women have to eat in a small table in a huddle. Though they ache in joints because of intense labor in a kitchen, they can't even st...
1 comment
Read more

Dormitory alone

     I throw myself on my bed being exhausted. I can hear my roommates under me talking loudly about food they are going to eat in home during weekends. Wishing the dormitory weekend menus to be good, I close my eyes for a short sleep until the first self study time begins.   Surprised with a large sound coming out from my alarm clock, I wake up. The total silence in a whole room makes me hard to believe that only 30 minutes have passed from the moment that the whole dormitory was full of sound of students being excited about going home. I consider sleeping for a whole self study for a moment, but I soon make up my mind and get out of the bed since I know dormitory teacher always look around the empty dormitory in order to check whether students cleaned their room before they leave.   I slowly put books in to study during the first self study time in my bag and go up to a cafeteria, my favorite place to study. A cafeteria which is usually full of students...
1 comment
Read more

there is no better place for me than KMLA..?

Q. there is no better place for me than KMLA. At this very moment, there is NOWHERE i'd rather be . Really..? A. I am sure that there is no better high schools than KMLA for me in Korea. I have always hated the atmosphere that 'force' students to study since I was very young and had always dreamed of dropping out of school where 'I' am never respected as I am. (I got to hate it since I was objected to major in arts by many adults though I had started drawing since I was 4 and was actually good at it just because 'I won't make much money if I major in design".) KMLA was a place where I actually 'escaped' from those forceful atmosphere as I thought I would be able to do what I want to do more freely than other places. I am completely satisfied with what I am doing in KMLA since I must not have been able to do activities like designing T-shirt, newspaper, and newsletter, taking photos, and running a play if I were in normal high schools. ...
1 comment
Read more