xiao qi 小奇

Kitten, borned in my house~

One hot afternoon, A big female cat jump into my room from window(why I know is female is because the visible big breast). After chasing her out, I check the 'drain' outside my room. (My room is at 1st floor, so there is a weird dead end 'drain' like hole for aircon discharge or planting i guess). Surprisingly, 4 kitten inside there! Problem is my hands cant reach out for them! so i modify a plastic to pull them out.

Now they living at the corridor 1st floor, my block. everyone feeding them except us. but they still like to play and sleep at our shoes rack, sometimes even in our shoes.

Only 3 kitten left, another one I cant find her anymore. very playful now. keep fighting and jumping although cant really walk properly.

Gf put a layer at the bottom of the shoes rack so they will nt fall into the gap. Then, start fighting for the space untill we put a shoes to separate the place into 2. Another one still playing with the slipper at the corridor.

Trying hard to get into the new bed!

What is this? she just curious about everything. a new born baby.

This is the weakest!
So innocent, but I still prefer dog. sorry...

My favourite little black, 小黑, very cute... and agressive when playing...

Playing with the tali...

Comfortably sitting on the new bed, separating with a shoes. neighbour with the little black.

Before putting a layer for them... see the weakest totally strandled by the sticks... stuck in there!

Sayang sayang, i dont like the cat everytime also so manja...

Trying to be funny har?

Another case of stuck in the sticks forest

Go dream la!

Sometimes, is good to day dream, especially during a boring, long, tiring experiment...

I'm have to admit that I'm a poor protein researcher, my salary is just enough to feed myself and a fish, sometimes cat and dog.

DSLR-digital single lens reflex camera will be the basic of my daydream usually, others than girl...

The Canon EF 70-200mm f/4L IS USM

The lens with a red ring and superb quality! Non IS version ~RM2985, IS version ~RM4685.

walao... where got money wor...
Canon BG-E5 battery grip for 450D

An accessory that can make my camera more solid and can tahan for longer time! no more worries about depletion of power! and better handling! ~RM575

Tamron AF 18-250mm f3.5-6.3 Di|| LD Aspherical IF Macro

All in one zoom lens, good for travel ma... near 14X zooming ability, why tamron? because cheaper than Canon lo... ~RM1,650

Canon EF 50mm f1.8 II

The cheapest lens and good value lens. its plastic feel is the disadvantage, need to be very careful, if drop, mean finish! However, the image quality from this lens wouldnt dissapoint anyone. ok la, I aim this prime lens la..~RM300!

Benro A350EX QuickLock Aluminium Tripod

Tripod ar tripod... It would be fun if I hav a tripod to play with! Why everything so expensive la...~RM320

Nissin Di622 speedlite

Flash gun! I gotta get you soon! just wait and see!

~RM400 3months ago, now seems like naik harga liao...-_-|||

ok, finish day dream, continue to transform bacteria! here I come!

nothing is imposibble!

I finished my secondary education in SMK Taman Johor Jaya (1) which is ranked number 2 problematic school during my time, and climbed up to number 1 after I'm grad because my super juniors just burned the security guard pondok.

I was actively involved in St. John Ambulance Malaysia then and enjoying school politics as Duty Secretary of SJAM division SMKTJJ1.

Form 6, I have to changed to another secondary school, SMK Permas Jaya, which is a better managed school, but the students there are just too good, they will walk within the line, everyone uniforms are so neat, the toilet is so clean, nobody smoke, no fire burning, no ppl trying to steal the TV from library and the staff and head master are just too good and fair! make me not used to it!

The SJAM here is like a dead organization. With 300 members in the list. Nobody appeared on Saturday Ko-K activity. No proper activity. No proper AGM. As SJAM members, we decided to help them up.

After several meeting and discussion, I found out many things that I thk is possible, is impossible in their mind, esp from Physics class.

This is opposite of my idea that we should try before we say it is not doable, even we failed, we still can learned something from the failure, of course, we need to plan properly.

After so many years, many ppl still worry this and that, aiya, just do it la when u still young, dont scare fail ma...

However, after so many years, I also kena this virus, injected into my mind liao.

Is time to think again and adjust myself.

IF it is not possible, and if I'm not trying hard enough, I will be regret and SJAM of my form 6 school will not be revived.

Just remind myself not to be too EGO, a little bit can la... haha!

Is time to really think about it.

Colleages

Working with molecular biology, you need imagination, unless you got a microscopic eyes that can zoom into molecules. Here are some tools we using.

A spectrometry, to check on the concentration of bacterial cells, protein, as well as DNA by the reflextion of light.

An Autoclaving machine, heat up to 121C for 15mins, 15bar, in order to sterilize culture media, keep our tools from contamination by microbes.

A weight balance, to measure weight for chemicals, media powder.

Water distiller, to produce clean water free of contaminated chemical like nuclease and protease.

A water bath machine, maintain desired temperature for incubation.

A Polymerase Chain Reaction machine, (PCR), amplification of genes.

A UV light detector, to visualize the results of electrophoresis for analysis.

Kubota centrifugal unit, to separate cells, DNAs, proteins @ desired temperature.

Agarose gel electrophoresis, to run the gel in order to know what u get is what u want.

Not what u want? run again then, u just wasted another 60minutes.

There are a lot other useful stuff for molecular biology such as SDS P.A.G.E for protein analysis, crystallization wells and others.

Mistakes

Sometimes, feeling hopeless and demotivated when things go too slow...

1. E. coli Rosetta-gami2(DE3) pET32-phaCcs

not stable, the expression level keep reducing after sometimes.

Revival of glycerol stock and verification ate most of my times.

Production is still OK. Some part in my mind is asking me to retransform the plasmid again.

However current priority is still crystallization(produce protein fast), and my target for crystallization is end of MAY!

2. Own stupid mistakes(thinking not diverse enough)

Purification in July

The purified PhaC protein seems not pure enough, so, I'm thinking to purify the elution again using the regenerated resin and I did.

However, the results of the SDS PAGE analysis is not good, because the PhaC protein cannot bind efficiently to the resin.

I thought is because of the Regenerated resins. Hence, I try new resin (mistake: I missed something very important here when troubleshooting).

For the 3rd purification, I used the elution from the 2nd purification. The result was the same. Binding of the protein is not efficient. Cannot figure out, think and think and think, and finally....I'm feeling stupid.

There are imidazole with concentration of 250mM in the elution that i used for 2nd and 3rd purification!

My large-scale protein production end here. no more protein. Now speeding for protein production again. I want xstal!

Research, Scientist and Laboratory

Is going to be 6 months soon... The number of time that I'm did my research in lab 414 molecular biology laboratory...

Basically, I like researcher's life, is always full of surprises and disappointments. Of course, very tiring and a very hard efforts are required. So far, There are 4 big stages of my project :

Cloning, Expression, Purification and the crystallization.

Cloning

Construct the desired DNA sequence into a suitable expression vector, and transform into a host, mostly E. coli.

In my case, I'm using pET32 expression vector from Novagen.

Transform into E. coli strain Rosetta-gami 2.

This stage done!

However, I'm trying another expression vector, pCold expression vector, and transform into classic of expression host, E. coli strain BL21 pLySs.

On the way!

Expression

To stimulate the bacterial host in order to over express (translate the gene seq on vector into protein) desired protein.

Optimizations were done, mainly on temperature. 15C, 25C, 30C, 37C. IPTG as inducer with concentration of 0.4mM.

Run SDS PAGE to determine the proteins.

Purification

Using TALON resin, going to try gel filtration soon. maybe can play ion exchange le? yohoo!

Others

6X His Protein Staining, Western Blotting, In vitro enzyme assay