meeting 16 -15 september

Present: (Aina, Shila, Arfa) -- arfa left at 1.42 pm

Time: 10 am - 3.30 pm

what do we gather
1) check od
time: 3.30 pm-10am
od: 2.080

2) measure the weight of the ethanol extract from the eppendorf tubes

A: 1.6825 - 1.0554 = 0.6271g
B: 1.7527 - 1.0500= 0.7027g

for A: 0.6271g were reconstitute with 1ml distilled water, vortex to dissolve
---> concentration: 0.6271g/ml

3) measure the zone of inhibition

*controls
- NaB 1.8
- E 1.3

*well

NaB, W, 10*3, 10*4, 10*5 ( NaB 1.6)

*disc

NaB, W, 10*3, 10*4, 10*5 ( NaB 1.7)


*well

NaB, W, 0, 10*1, 10*2 ( NaB 1.8)

*disc

NaB, W, 0, 10*1, 10*2 ( NaB 1.7)

*well

W, NaB, E, PJ ( NaB 1.8)

*well

W, NaB, E, PJ ( NaB 1.5)


4) do agar disc and well diffusion ( incubate at 1.45 pm )

Well :

- NaB, W, PJ
- NaB, W, E
-NaB, W, HW

Disc: ( 25 ul 1st, let it dry, another 25 ul added )

- NaB, W, PJ
- NaB, W, E
-NaB, W, HW

5) innoculate different concentration of PJ , Ethanol and control (incubate at 3.15pm )

Pure juice

A- 5ml broth & 50ul e.coli & 200 ul extract
B-5ml broth & 50ul e.coli & 300 ul extract
C- 5ml broth & 50ul e.coli & 400 ul extract

ethanol

1-5ml broth & 50ul e.coli & 200 ul extract
2-5ml broth & 50ul e.coli & 300 ul extract
3-5ml broth & 50ul e.coli & 400 ul extract

control

5ml broth & 50 ul e.coli

6) innoculate 10ml broth and 100 ul e.coli (I) (incubate at 3.15 pm)


7) grind ginger using food processor and measure 0.1076g into agar well with NaB and W.
(incubate at 2.30 pm )


next plan of action:

check OD for the e.coli and the different concentration of PJ and ethanol
check zone of inhibition
do serial dilution

ohh nooooo

FYP had been hectic these few days.

As of today, early in the morning I got a call from Aliza and sort of scolding..


*ring ring*

shl: hello?
ali: hello. shila! what time is u all coming?? u said 9.30?!! now what time already you know?? now 9.45 already! where are you??
shl: +aina what time we reach there??+ Ohh ermm.. we at woodlands already..
ali: HUH??!! WHERE??
shl: we at woodlands interchange! +almost shouting+cos she like cannot hear me. =.=
ali: WHAT TIME YOU ALL COMING?
shl: +aina what time??+ ermm. around 10.
ali: WHAT? 10.30???
shl: no.. 10!!!!
ali: oh ok.
shl: ok.

there was no bye or whatever. =.=
and at the point of time, the 903 bus just have to come rather late. or maybe I think becuase we were too anxious already larh. grrr. so scary can?!

when we reach the lab at 10am, we were so scared don’t know what to say. but luckily she didn't scold or something. and i just realize we didn't say sorry or what. brrrr.. and suddenly she stayed in the lab with us the whole time today. REALLY!

Anyway, when other facci (the guy with a very low scary voice) came in our lab, I told aina that we should be glad to have Aliza instead of other PI?? right arfa??

anyway I’m trying to be strong when facing Aliza these few days. and also the lack of manpower....+speechless+ I’m not blaming anyone but how can even ARFA always got SOMETHING on and have to go back early when he is ALWAYS late to turn up early in the morning?? Today he said he was otw when me and Aina were in the MRT. we reach the lab but he is still not there. He arrived about 1hr later larh. Pei, I think he put the eyelash extra long and super mega subtle that’s why it looks natural. that's why need 2hrs to MAKEUP. Walaowei.

ANW! Pei I heard you are not feeling well today. hope you feel better! Come back soon yo!

meeting 15 - 14 september

Time: 2.30pm-3.30pm (Aina, Shila, Arfa)

What do we gather?
1) Do LB agar
--> 2 500ml bottles
--> 12.5g LB + 7.5g agar = 500ml distilled water
--> total made: 1000ml

2) Inoculate bacteria
--> H

3) Put ethanol extract in vacuum concentrator.
--> We decided not to put it in the rotary evaporator.
--> Aliza suggest we put it in the vacuum concentrator to spin dry the extract. (we got use before last sem)
--> separate the extract into 2 microtubes.
--> put in vacuum concentrator overnight at 60dc. (3.30pm-10am)
--> empty microtubes A- 1.0554g & B- 1.0550g


Next plant of action:
- check OD
- pour plate
- do agar well and disk diffusion
- check ethanol in the vacuum concentrator


*last but not least,

meeting 14 - 11 september

Time: 9.30-10.30am

What do we gather?
1. Bacteria on the plates are confluent. There is no isolated colonies found even for the 10^7.
2. Plate A-10^7 is missing. Can'r be found anywhere. (might have been misplaced)
3. Ironically there is absolutely no bacteria growth on B-10^7 but there is confluent growth on C-10^7. That should no be the case.
4. There is something wrong with the pattern of the bacteria growth. It is only on one side or not evenly lawned.
5. Bacteria growth on all the plates are not as we wanted.

Possible reasons why it happen:
- Plates were not lawned/spreaded properly
- There might be a mixed up in the spreading of the dilution.
- Bacteria in the tubes might have growth by the time we did the spread plate as we waited from 1+pm all the way to 4+pm to spread and left the serial dilution solutions all in the BSC. (which might lead to the confluent growth)

Corrective measures/next plan of action:
- We need to do the serial dilution again. (this time, do not rush when spreading and bacteria are spread immediately after doing serial dilution. no waiting. time planning people)


*note: I didnt take pictures of the serial dilution plates cos it look awfully disgusting and it just hurt me just to take pictures of our failures.

MANY SOBSSSSSSSSSS!

meeting 13 - 10 september

Time: 8.30 to 5.30 pm

1) Check OD for the different concentration of pure juice and hot water

Pure juice

A- 5ml broth & 50ul e.coli & 200 ul extract ------------- 2.0049
B-5ml broth & 50ul e.coli & 300 ul extract ------------- 2.0873
C- 5ml broth & 50ul e.coli & 400 ul extract ------------- 2.3452

Hot water

1-5ml broth & 50ul e.coli & 200 ul extract ------------- 1.864
2-5ml broth & 50ul e.coli & 300 ul extract ------------- 1.668
3-5ml broth & 50ul e.coli & 400 ul extract ------------- 2.322

control

5ml broth & 50 ul e.coli ------------------ 1.868

G --------------- 1.994

2) make LB agar

12.5 g LB + 7.5g agar --> 500 ml water ( 20 plates )
25g LB + 15g agar --> 1000 ml water ( 40 plates )


3) serial dilution for the pure juice, hot water and control

- measure 900 ul of saline into 100 ul of bacteria into the A, B, C, 1, 2,3 and Control. eg:

A

10^1
10^2
10^3
10^4
10^5
10^6
10^7

from there, only 10^4 until 10^7 for each set were plated and incubate at 5.30 pm

meeting 12 - 9 septemebr

Time: 9am-3 pm

1) check OD
time: 3pm - 9.15am
od: 1.958

2) check plates
- no zone of inhibition for all the plates except for the NaB.

DISK DIFFUSION

controls, original, 10^1, 10^2

controls, 10^3, 10^4, 10^5
controls, ethanol, pure juice

WELL DIFFUSION

controls, original, 10^1, 10^2

controls, 10^3, 10^4, 10^5

controls, ethanol, pure juice

3) do saline : 250 ml of water with 2.25g of NaCl and autoclave at 12

4) do a bigger paper disk (using the cap of a green falcon tube) and autoclave at 12

5) innoculate different concentration of pure juice and hot water.

Pure juice

A- 5ml broth & 50ul e.coli & 200 ul extract
B-5ml broth & 50ul e.coli & 300 ul extract
C- 5ml broth & 50ul e.coli & 400 ul extract

Hot water

1-5ml broth & 50ul e.coli & 200 ul extract
2-5ml broth & 50ul e.coli & 300 ul extract
3-5ml broth & 50ul e.coli & 400 ul extract

control

5ml broth & 50 ul e.coli

6) innoculate e.coli, 100ul of e.coli in 10 ml of broth (G)

7) do a set of all the controls using well diffusion method

- NaB
- Ethanol
- Water
- Saline

**** all the innoculation and controls were incubate at 2.30 pm

meeting 11 - 8 september

Time: 8.30am - 4pm

1. check od
time: 3.18pm - 8.50am
od: 1.943

2. rotary evaporator
hot water: 72mbar, vapor temp - 41dc, 80rpm, water bath - 95dc
ethanol: 175mbar, vapor temp - 42dc, 70rpm, water bath - 65dc
chiller: 5dc

3. hot water extract
extract with flask - 93.440g
extract diluted with 1ml distilled water
empty flask - 93.108g
extract - 0.332g
concentration - 0.332g/ml

4. serial dilution of hot water extract
100ul into 900ul of distilled water
do till 10^5 dilution

5. disk and wel diffusion method

disk: 30ul, well: 50ul
time:
well - 3.48pm - 9.15am
disk - 3.56pm - 9.15am

6. innoculation
time: 3pm - 9.15am

meeting 10 - 7 september

Time: 3pm - 3.30pm

1. Innoculation
Time: 3.18pm - 8.50am

2. checking control plate
both have no bacteria growth

meeting 9 - 3 september

Time: arfa (9am - 1pm), 11.30am - 4pm

1. check od
time: 3.15pm - 9.15am
od: 1.91

2. control plate
- spread plate for hot water and ethanol extraction to test for sterility
- time: 11.26am - 3.18pm (7 sep)

Pics & Vids on 8th Sep~

Hello all! I'm here to post the pictures taken on 8th Sep, yesterday, and also the result of the DISK and WELL diffusion that we did yesterday too.

Have fun and enjoy looking at the pics especially ARFA GRAND MAGIX SHOW!

THANK YOUU!

water henna on aina's hand -done by shl

baka arfa kept laughing alone.

and laughing again

and aina end up laughing with arfa! baka.

peiyi long hair

how to weigh a round bottomed flask?

so neat and tidy

rotary evaporator

Rotary Evaporator




ARfa GRand Magix Show

meeting 8 - 2 september

Time: 10am - 1pm, 3.15pm (Arfa)

1. Result of serial dilution
10^7 = 63 colonies
CFU = 6.3 x 10^9








2. Check OD
time: 4pm - 10.20am
e.coli: 1.911

3. Innoculation
time: 3.15pm - 9.15am

4. Extraction of ginger
ethanol and hot water extraction filtered using vaccuum and pump.

meeting 7 - 1 septemeber

Time: 9.30am - 4pm

1. Check OD
time: 4pm - 9.30am
e.coli = 1.508

2. growth curve
using one colony from streak plate and put into broth.
incubate at 10.50am, 37dc and 100rpm.
measure every 1 hour +/- to obtain 6 points to plot the graph.
11.50am - 0
1ml was mistakenly thrown away by lala
1.18pm - 0.183
2.18pm - 0.669
3.18pm - 1.378
3.30pm - 1.438
4pm - 1.656
[draw the graph yourself.]

3. preparation of ginger
washed > peeled > washed > dried > cut > blended > 1,2,3
1 - hot water
- 200g soaked in 100ml of hot water for 24 hours (1.30pm - 1.30pm next day)
- resultant juice extracted, stored in bottle and placed in box and ready to use for testing.
2 - 95% ethanol
- 200g soaked in 100ml of ethanol for 24 hours (1.30pm - 1.30pm next day)
- extract obtained, stored in bottle and placed in box for further processing.
3 - cloth
- 200g.
- pulp were separated using cloth
- pure juice was extracted and stored in dark bottle at 4dc.










4. serial dilution
time: 4pm - 10.19am

5. innoculation
time: 4pm - 10.19am