Preparation of chemicals for BOD

4 solution required to be added into the sample:

To make 1L solution:

• Phosphate buffer solution

8.5g of potassium dihydrogen phosphate (KH2PO4)

33.4g of sodium hydrogen phosphate heptahydate (NA2HPO4.7H20)

1.7g of Ammonium chloride (NH4Cl)

21.75g of Dipotassium hydrogen phosphate (K2HPO4)

*add all up and top up with distilled water till the level

• Magnesium sulphate

22.5g of MgSO4 + distilled water

• Calcium chloride

27.5g of CaCl2 + distilled water

• Ferric chloride

0.25g + distilled water

KLCC

looks yummy, but sorry to say, its tasteless. we have tried few of it.

macaroons...not as nice as at WhiskBar

this is yummy for sure, my mee kicap.wanted to eat this for quite a long time already

mom with her delicious mee kari, her fav dish.

view from above

celebration of Christmas, a month to go. not for me, but ull get to see lots of nice decorations in most of the shopping mall here in KL.

Short Film Festival

Unisel, starts from 3-7pm.

New lab & I hate the environment

Bench work to prepare my standards

My bench work for IC Anion 9 elements

what can i say? Ive moved to my new lab but I must say that I hate it so much..Nobody disturb us and make a lot of noise when we r doing experiment at our old lab, nobody will ask me for gloves, for standards, for what ever it is...

Purified DNA round 4

the purified band for R1 and R2

Purified DNA --> Test for the DNA purified -->;Prepare for PCR --> PCR --> Run gel electrophoresis = Get the DNA band

Chemical Kinetics - Reaction Rates

Im trying to understand exactly what does these orders means..damn, who ask me not to study real hard during my diploma???? now what? i can still refer to the internet and reference book off course...

well, found 2 web pages that is useful

http://www.chem.arizona.edu/~salzmanr/480a/480ants/kinintro/kinintro.html

http://www-teach.ch.cam.ac.uk/teach/IA/KCR_full.pdf

Another set of primer

Ive ordered again, new set of primes. The old one has finished..I should ordered it last week, Im unaware that the PCR has finished.

Here are my Primers, i refer to O' Sullivan's and Muyzer's paper. My 1st time ordering primer attached with GC clamp.

Synthesis ID: 1051372
Primer Name: 357F_GC
Sequence (5'-3'): CGC CCG CCG CGC GCG GCG GGC GGG GCG GGG GCA CGG GGG GCC TAC GGG AGG CAG CAG
Scale: 100 nmole
Purity: PCR Grade

Synthesis ID: 1051373
Primer Name: 518R
Sequence (5'-3'): ATT ACC GCG GCT GCT GG
Scale: 100 nmole
Purity: PCR Grade

PCR round X

before purifying DNA for R1

before purifying DNA for R2

X-shows infinity...hahah not sure already how many times im doing it..luckily its all in my log book..

Ive to do it all over again since the purified samples prepared previously have been used for DGGE but unfortunately, no results. Still figuring out whats the problem. ive asked some experts, they were also not sure and aware of my problems..

1-Preparation of PCR samples, 25microlitre (mixture of DNA; R1 and R2 + nuclease free water + primer 357F-GC and 518R+Green Master Mix)

2-Run for PCR, programmed A at CDDR lab, machine 1 (near the sink) and for about 6 hours, starts at almost 10.30am, finished at about 4pm.

3-Prepare my gels at Brain lab, so i need 2 large sets of agarose gel. (mixture of 6microli of etbr + 140ml of 1X TAE buffer + 1.8g of agarose powder)

4-Mix up the DNA ladder, 100bp and orange loading dye. (mixture of 5 and 2 microlit respectively)

5- Load all my samples, Gel D is for R1 and gel E is for R2. 15 wells all together. 14 samples and 1 well for control.

Back pain

Quite frustrating with what's happening to me now..

Im suffering from back pain, starts from my waist to my thigh...it hurt so much..

I tried to sit straight up, not to overly bend my body and be very careful with what im doing now..

i guess its all because of my last year's work...I lift up heavy loads; my wastewater samples, moving boxes here and there; moving house, moving labs..

And its killing me now, hope its gonna heal, i know it will takes a long time...

Im using counterpain, im applying cold as well, i just bought the reusable ice pack from guardian, cost me about RM23.90.

Lab

today is quite boring, normal routine..

Prepare samples for PCR, each sample i prepare about 14 pcr tubes, about 25microlit each.., quite tiring.

Then, wait for the samples to run for about 6 hrs, again boring

Prepare 2 agarose gel at brain lab for each sample

Then load 5microlit of sample into each well (28 wells all together)

Mix up the loading dye and 100bp DNA ladder for both gel

Now, waiting for the gel electrophoresis for another 40minutes...damn, tired of waiting.!

Well, here are the results for both samples; R1 and R2.

14 samples of PCR, only tube 1 shows no DNA sample, can just discard it. No need to do further purification for tube 1. The rest, from tube 2 to 14 was just great! lovely bands..

For R2, seems that all PCR tubes prepared contains DNA, based on the bands appeared. Lovely.

Alex's birthday celebration

Alex's pirate cake

The decoration by Alex's mom. Theme: Pirates of Caribbean

Me with the pirate moustache

Our pirate ring, isn't it cute?

Me with Alex's mom, Ida

Alex is turning 3 this year! I went to her house with sis at Tiara Kelana Kondo. It was my 1st time visiting her here. We have pirate cake, fried rice, Domino's pizza, drinks and etc.

Im at FINAS

The hall, sis said this place was also use when they want to shoot for scene in film or drama

the tentative book, nice one..very detail.

Me with clapper board at FINAS

The open area at FINAS building

Im with the stars...P.RAMLEE, A.R TOMPEL..

Its my 1st time, we reached there at around 11.30, just the right time as the session 1 has finished...damn, the next session starts at 2pm. so we waited till 2pm just to watch these series of international Short stories...

Starts with story from Japan, Germany, France, Malaysia and Korea...but what i like most is the Germany, Title: Interview. Thriller. U can even imagine how the story going to ends but thats it... full of mystery.

My new workstation

my new workstation at the FCE, pg room.

ABC

ABC at Pelita Nasi Kandar, Section 9, Shah Alam. RM 4.00

Noodle station

Luncheon yesterday with cm and nik @ the noodle station SACC Mall. I ate this, some sort of kueh teow with beef pastrami and iced peach tea..

Lovely cousin

Im with Husna, met her yesterday..Will be meeting her at least once in 2 weeks time...so cute and adorable...hehehe love her so much and keep on missing her every seconds...

Lunch as a whole

Lunch at Zam Zam Oasis 9, Shah Alam

DGGE Gel

Today I have to prepare chemicals for DGGE gel for about 3 times. quite a lot n tiring.

1-I tried to fill up the gel into the set up, but it's not working.

2-I tried it with Ida, somethings not right too, so I have to do again. The same thing happen.

3-A little bit of patience required here. huhu so i prepare all over again, 0 and 100% of stock solution and for Low and high density..

so many chemicals involved, the addition of 50X TAE Buffer, Formamide, Acrylamide bis, Urea and distilled water. Dcode dye solution also required, APS...

Preparation of Buffer for DGGE

Adding up distilled water into my buffer container

U'll need about 9 litres of 1X TAE buffer to run for DGGE.

So I use the 50X TAE buffer prepared previously.

To make a 9 litres 1X TAE Buffer, 180mL of 50X TAE buffer is required.

Prepare your buffer container, add up 720mL of distilled water and top up with 180mL of 1X TAE Buffer.

Done.

Cont on gel electrophoresis

band appeared for purified DNA after PCR on gel for R1 and R2

On the left, R1, on the right R2. only 3 microlit of purified DNA used for all bands appeared here

DGGE gel, not working this morning. No idea y. so blur..n a bit frustrated..

damn, tension...supposed to be good, dont know what went wrong.

Pose while doing experiment

cont DNA purification

the set of DNA purification kit

purified DNA for both R1 and R2

Adding Membrane wash solution into the minicolumn

Purification process of DNA - round 2

Discard fluid in Collection tube

After centrifuge