I've been in my new lab for one month. I did kinds of pre-works: making batches of solutions and buffers, and, if you wonder more, that was, washed piles of flasks, and them bake some of them at 200 centigrade for 6 hours; then filter the solution one by one and thus made them eligible for RNA-related experiments.
As a result, I filtered 15+ solutions, made about 20+ solutions, and 20+ buffers. My firstly-empty rack was full now.
Such work was not unfamiliar, but there were always kinds of new "single lab specific tips" waiting for me to know. No no aseptic rooms, no ultraviolet lights, no much of kits (in fact they only have one gel extracting kit), even no lap spoons, (the last one was for avoiding the cross-contaminant; actually I wonder if they will use alcohol burner when dealing with microbes... o__O ) They (soon the word "they" would become "we") make everything by themselves (ourselves).
"We buy nothing but only chemicals."
That was very true. Firstly, I thought at least there should have some essential buffers and reagents. But later I knew I was wrong. We have to make PCR buffers, dNTP mixture, the loading dye.
After done all of these liquids, I did my first PCR on September 28.
In my previous lab, we didn't use EB and used GreenView instead. Here we use EB (ethidium bromide) to dye DNA. Thus it could be the first time for me to contact EB. When I dyed the gel in EB-water, a senior guy here told me something important:
"Remember to change your gloves every 10 minutes if you're working with EB. EB will be able to go through your glove and then go into your body after contacting of 10 minutes."
Fine. I often change my gloves. No worries. I told him.
"Yeah, I know. Anyway..."
.
.
.
"I'm not joking!"
He said with a serious joking way of speaking and a funny emotion.
Wah! LOOL!
But I saw nothing but the marker DNA on my gel later.
My PCR failed? What happened? Because I did a positive control, we confirmed the problem was about the Taq polymerase soon.
"Hmm... I've ever doubted that batch of Taq. We need to re-purify some."
We need to re-purify some.
What? Not just adding the powder of Taq into its buffer? Re-purify?
I knew already that there's no ready-to-be-used Taq liquid, and I firstly thought they buy the power of Taq.
"No. Actually, in this lab, we start from zero. We make everything, that including Taq, Pwo, and T7. Here is such a lab where you need to do EVERYTHING."
I see, I see. That means, I can train myself very much, huh?
I'm not joking!
Hey, be more positive! Although it's a bit troublesome and time-consuming, I haven't seen the procedure of purifying some Taq polymerases and then use them in PCR. Nice chance!
Okies! I'm waiting for looking at the whole purifying of Taq, maybe Pwo as well! :3
As a result, I filtered 15+ solutions, made about 20+ solutions, and 20+ buffers. My firstly-empty rack was full now.
Such work was not unfamiliar, but there were always kinds of new "single lab specific tips" waiting for me to know. No no aseptic rooms, no ultraviolet lights, no much of kits (in fact they only have one gel extracting kit), even no lap spoons, (the last one was for avoiding the cross-contaminant; actually I wonder if they will use alcohol burner when dealing with microbes... o__O ) They (soon the word "they" would become "we") make everything by themselves (ourselves).
"We buy nothing but only chemicals."
That was very true. Firstly, I thought at least there should have some essential buffers and reagents. But later I knew I was wrong. We have to make PCR buffers, dNTP mixture, the loading dye.
After done all of these liquids, I did my first PCR on September 28.
In my previous lab, we didn't use EB and used GreenView instead. Here we use EB (ethidium bromide) to dye DNA. Thus it could be the first time for me to contact EB. When I dyed the gel in EB-water, a senior guy here told me something important:
"Remember to change your gloves every 10 minutes if you're working with EB. EB will be able to go through your glove and then go into your body after contacting of 10 minutes."
Fine. I often change my gloves. No worries. I told him.
"Yeah, I know. Anyway..."
.
.
.
"I'm not joking!"
He said with a serious joking way of speaking and a funny emotion.
Wah! LOOL!
But I saw nothing but the marker DNA on my gel later.
My PCR failed? What happened? Because I did a positive control, we confirmed the problem was about the Taq polymerase soon.
"Hmm... I've ever doubted that batch of Taq. We need to re-purify some."
We need to re-purify some.
What? Not just adding the powder of Taq into its buffer? Re-purify?
I knew already that there's no ready-to-be-used Taq liquid, and I firstly thought they buy the power of Taq.
"No. Actually, in this lab, we start from zero. We make everything, that including Taq, Pwo, and T7. Here is such a lab where you need to do EVERYTHING."
I see, I see. That means, I can train myself very much, huh?
I'm not joking!
Hey, be more positive! Although it's a bit troublesome and time-consuming, I haven't seen the procedure of purifying some Taq polymerases and then use them in PCR. Nice chance!
Okies! I'm waiting for looking at the whole purifying of Taq, maybe Pwo as well! :3