meeting 30 - 3 november

time: 4.30pm - 6pm
all present except ainah

- tlc run 2
did tlc run 2 to comfirm the result done at tlc run 1.

- process:






- result:

meeting 29 - 30 october

time:
(aina, fadh, shila):
arfa left early:
py absent

1. make trolox solution
0.0015g into 50ul h20

2. label the plate
+, -, L, G

3. put ?ul of each solvent onto TLC plate

4. pour toluene and ethyl ecetate into a beaker and cover with aluminium foil. fill until half of the pencil marking on the tlc plate.

5. run the plate until almost near the top. draw a line.

6. air dry the plate

7. immerse in 5% ethanolic sulphuric acid and then 1% ethanolic vanillin.

8. heat the tlc plate on hotplate until color spot appear.

Result:




please interpret together.

meeting 28 - 28 october

time: someone please tell the time at tagboard.
attendance: all present except py.

Discussion with aliza before running TLC.
Someone please update this.

meeting 27 - 20 october

time: 4.30pm - 6pm
all present

Suggestions of failure:
- wrong experimentation design
- extracting failure
- did not use ginger (pure) immediately
- combination would have cause a better effect

Lime: results shown

Why lime gave the results?
- Properties (Physical/Chemical)

Does lime acidity (pH) has any effect on the zone of inhibition?
If yes, how to prove?

- What causes the inhibition in lime? The active ingredients?

- No consistency in the results. Need to state how much is the different. Double?

* Need to tabulate results!

What cause the different results in the runs?
- storage of extract
- amount
- different people measure (not valid!)
- time of measurement

- Why S.epidymidis? Why use e.coli?

Things need to be done:

- Send Aliza TLC protocol
- Next lab session: Monday, 26th
- Report

1. result of run 3:

Lpg - 1.1cm
nab - 0.9cm


nab - 0.8cm
L - 1.1cm


nab - 2cm
Lpg - 1.5cm


nab - 2.2cm
L - 1.6cm
Leg - 1.1cm

2. summary of all 3 runs:


3. reasons for inconsistency in the result:

meeting 26 - 16 october

time: waiting for someone to tell me
present: waiting for someone to tell me

s.epi serial dilution plates result
did anyone count the cfu???

meeting 25 - 15 october

10.30 – 11. 15 am (aina and fadh)
4.30- 5.30 pm (aina and fadh)

Check 2nd S.epi OD from 6pm (yesterday) to 10.45am (today). (BS) --- 1.625

Do inoculation for 3rd S.epi (CS) from a S.epi streak plate (100ul in 10ml broth) at 11am and check OD at 4.45pm. --- 0.108

Do serial dilution from 3rd S.epi. and do spread plate and incubate at 5.30pm.

meeting 24 - 14 october

time: 11.30pm - 5pm (fadh), 4.30pm - 6pm (aina, shila, peiyi)

1. growth curve of s.epi
innoculate at: 12pm in incubator shaker, 100rpm, 37dc (15ml)
1pm - 0.084 (14ml)
2pm - 0.104 (13ml)
3pm - 0.142 (12ml)
4pm - 0.202 (11ml)
4.45pm - 0.212 (10ml)
5.15pm - 0.408 (9ml)
6pm - 0.814 (8ml)
100ul for innoculation.

2. innoculation
100ul into 10ml broth

3. LB agar and broth

prepare, autoclave and pour plate

total: 47 plates and 750ml broth

meeting 23 - 13 october

time: 4.20pm - 5.45pm (all present except aina)

1. check od (k)

od: 1.895

time: 5pm till 9.30am

2. prepare LB broth and agar

requested already for 50g LB + 19g agar

our plan is to make 40 plates and 750ml broth

broth: 18.75g LB into 750ml water

agar: 25g LB + 15g agar into 1000ml water

3. run 3

time: 5.40pm - 11am

disk (30ul) and well (50ul)

tapwater, Nab, lime, lime mix pure ginger, lime mix ethanolic ginger

meeting 22 - 12 october

time: 4.30pm - 5pm

present: all except arfa

1. innoculation

100ul e.coli from G into 10ml of broth

time: 5pm till 9.30am

meeting 21 - 9 october

time : 1.30 pm -2.30 pm

time: from 6.00 pm (thurs) -- 1.30 pm (Friday) -- 19 hours +

Use your super incredible eye power to see! HEHHEHE!

hahahaha!!!!!!!!!!!!!!!!!!!!!!!!!!!! i know la ur HP so "GOOD" BAKA haha

k so..

DISK

1st plate :

NaB -- 1.9cm

L -- 0.8cm

2nd plate :

NaB-- 1.5cm

WELL

1st plate :

NaB--2.5cm

L&G--1.7cm

L--1.9cm

2nd plate:

NaB--1.9cm

L&G--2.0cm




TW: tap water

DW: distilled water
L: lime
LG: lime + pure extracts ginger
S: saline
+: NaB



DS: (written wrongly, actually is DW) distilled water
E: lime + ethanolic ginger
SB: NaB (bottom right)









Interpret yourself in your own logbook.

meeting 20 - 8 october

? - ? (arfa)
5pm - 6.15pm (la, na, pei)

1. check od for s.epi
time: 6pm - 12pm
od: 1.651

2. check plates for antimicrobial testing
disk:
nab: 0.8cm
distilled water: none
lime: 1cm
lime+ginger(ethanolic): 0cm

well:
nab: 1.6cm
distilled water: 1.1cm
lime: 1.7cm
lime+ginger(ethanolic): 1.1cm
- do the intrpretation yourself!




3. run 2
mixing of 500ul of pure juice of ginger and 500ul of lime to make 1ml.

time: 6pm - 1.30pm

meeting 19 - 7 october

10am - 10.30 am (fadh)

4.30pm - 6pm (all except fa)

1. check od (J)

time: 5pm -10.30am

od: 1.632

2. innoculation for s.epi

- take one colony from streak plate

time: 6pm - 12pm

3. antimicrobial testing - ethanolic extract of ginger+lime, lime

mixing of 500ul of ethanolic ginger and 500ul of lime to make 1ml.

reconstitute ethanolic ginger: 1ml distilled water into 0.7027g of extract

disk: nab, distilled water as negative control, lime, ginger+lime (30ul into each disk)

well: nab, distilled water as negative control, lime, ginger+lime (50ul into each well)

time: 6pm - 11.45am

meeting 18 - 6 october

Time: 3pm- 5pm
Present: Aina
Came later after school: Pei Yi, Shila, Fadhilah
Absent: Arfa

1. processing lime
5 limes of 370g are being extracted and the pure juice is being measured with 75ml and were keep in a glass bottle and stored in a cold room. The seeds and skin of the lime were stored in falcon tubes. Inoculate at 5pm.

2. innoculation (J)
time: 5pm - 10.30am

meeting 17 - 16 september

Time: 9.30 am – 4 pm
Present: Aina, Shila, Arfa
Absent: Pei Yi. Fadhilah

1. Check OD for E.Coli and the different concentration.

Pure Juice
A - 2.454
B - 1.110
C - 1.148

Ethanol
(1) - 1.197
(2) - 1.524
(3) - 2.675

Control (e.coli) - 1.394
I - 2.164

Time: 2.15pm - 9.45am (18+ hours)

2. Do serial dilution of Pure Juice, Ethanol and Control to 10^7.
Spread plate each set from 10^4 till 10^7.

Next plan of action is to check the serial dilution plates.

___________________________________________________________________
Result of all the serial dilution plates:





A4 – lot of colonies, uncountable.
A5 – confluent, not spreaded properly, lesser colonies than A4.
A6 – 36 colonies, lesser than A5.
A7 – clear of colonies.
the colonies decreased as the concentration decreased. confluent may be due to high od of 2.454.







B4 – confluent, uncountable
B5 – confluent, 100 colonies can be counted, not spreaded properly.
B6 – 32 colonies. lesser than B5.
B7 – 6 colonies. lesser than B6.
the colonies decreased as the concentration decreased. confluent may be due to high od of 1.110







C4 – lot of colonies, uncountable.
C5 – 300+ colonies
C6 – not spreaded properly
C7 – not spreaded properly
the colonies decreased as the concentration decreased. confluent may be due to high od of 1.148.

comparing A7, B7 and C7, as the extract increased, the bacteria growth increased too. this shows that pure juice of ginger does not have antimicrobial effect against e.coli. the optimal amount to inhibit bacteria could be around 100ul of pure juice extract.






14 – confluent, uncountable
15 – confluent, uncountable
16 – confluent, uncountable, not spreaded properly
17 – not spreaded properly, lesser than 17.
confluent could be due to not well spreaded. there might be contamination that affect the concentration.







24 – confluent
25 – confluent, not spreaded properly
26 – confluent, not spreaded properly, lesser than 25
27 – confluent, not spreaded properly, lesser than 26
confluent may be due to not well spreaded or contamination that affect the concentration.

34 – confluent
35 – confluent
36 – confluent
37 – confluent
too confluent could be due to high od of 2.675. there might be contamination that affect the concentration.

comparing 17, 27 and 37, 27 has lesser bacteria growth than 17. 37 has the most bacteria growth. the optimal amount of inhibiting bacteria could be around 300ml of ethanolic extract.

Cn4 – confluent
Cn5 – confluent
Cn6 – confluent, not spreaded properly
Cn7 – confluent
there might be contamination that affect the concentration. plates were not well spreaded. hence this is not a good control.

All the plates result don't tally with the od and therefore this serial dilution is not valid.