the unluckiest day
The unlukiest day...
I was very busy (or should I say lazy to update this blog) these few days...
To summarize everything:
Day 17 - repeat
When I did sequencing, ZD was not there. So I asked Wang Li and he told me that I only need to do 2 out of 10 samples so I only did 2. When ZD came back he told me that I need to do all 10 because this is the first sequencing as no one ever try to clone the gene before so I have to repeat the whole procedures for 8 more samples. At the end of the day I need to prepare bacteria culture for Midi preps the next day.
Day 18 - Linearization
After the midi-prep, I did a test digestion to determine the orientation of the insert.
Since I am going to do in situ hybridization, I need to linearize the plasmid that are going to be used as a template for constructing the probe. It's basically only digestion with enzymes. I just left those in the incubator overnight
Lab meeting - weekends
The presenter was Wang Li and dr Yi. But they were talking about our Lab secret experiments so I could not reveal those in this blog. After the meeting, I went back to the lab to take out the linearized plasmid and then help Ni to fill in tips. She paid me 10 cents for every box....
Day 19 - running gel
ZD asked me to run gel to determine whether the plasmid had been completely linearized. Unfortunately only 1 has been linearized. ZD asked me to make another linearization and somehow after 3 hours, I still get the same result... So in the end, I resort to gel extraction. I just load all my samples to the gel and run the gel for about 1 hours. After that I cut the gel and freeze it as I had no time to extract the gel
Day 20 - the unluckiest day
After gel extraction, ZD told me to purified the gel. However, he forgot to tell me that I should use DEPC water instead instead of TE buffer as the template need to be RNAse free. So he told me I need to repeat everything from linearization. Luckily I still have some plasmids left.
Today, SQ brought the pillow that I asked her to buy from IKEA. I had been suffering from a severe pain on the upper part of my thigh everyday after lab as a result for sitting for long hours. Then ZD told me that pillows are forbidden in the lab to avoid contamination as the fabric is absobant and hard to clean. So I wrapped the pillow in thin polyethene layer. Everyone was just giggle to see the pillow deflatten and made a farting noise everytime I sat on it.
I was very busy (or should I say lazy to update this blog) these few days...
To summarize everything:
Day 17 - repeat
When I did sequencing, ZD was not there. So I asked Wang Li and he told me that I only need to do 2 out of 10 samples so I only did 2. When ZD came back he told me that I need to do all 10 because this is the first sequencing as no one ever try to clone the gene before so I have to repeat the whole procedures for 8 more samples. At the end of the day I need to prepare bacteria culture for Midi preps the next day.
Day 18 - Linearization
After the midi-prep, I did a test digestion to determine the orientation of the insert.
Since I am going to do in situ hybridization, I need to linearize the plasmid that are going to be used as a template for constructing the probe. It's basically only digestion with enzymes. I just left those in the incubator overnight
Lab meeting - weekends
The presenter was Wang Li and dr Yi. But they were talking about our Lab secret experiments so I could not reveal those in this blog. After the meeting, I went back to the lab to take out the linearized plasmid and then help Ni to fill in tips. She paid me 10 cents for every box....
Day 19 - running gel
ZD asked me to run gel to determine whether the plasmid had been completely linearized. Unfortunately only 1 has been linearized. ZD asked me to make another linearization and somehow after 3 hours, I still get the same result... So in the end, I resort to gel extraction. I just load all my samples to the gel and run the gel for about 1 hours. After that I cut the gel and freeze it as I had no time to extract the gel
Day 20 - the unluckiest day
After gel extraction, ZD told me to purified the gel. However, he forgot to tell me that I should use DEPC water instead instead of TE buffer as the template need to be RNAse free. So he told me I need to repeat everything from linearization. Luckily I still have some plasmids left.
Today, SQ brought the pillow that I asked her to buy from IKEA. I had been suffering from a severe pain on the upper part of my thigh everyday after lab as a result for sitting for long hours. Then ZD told me that pillows are forbidden in the lab to avoid contamination as the fabric is absobant and hard to clean. So I wrapped the pillow in thin polyethene layer. Everyone was just giggle to see the pillow deflatten and made a farting noise everytime I sat on it.
posted by Melinda Leonie @ 1:17 AM
0 Comments:
Post a Comment
Subscribe to Post Comments [Atom]
<< Home