March 31 – April 14
Plasmid resource mining
- sampling
- strain isolation and purification
- Species and Genus Identification of Bacterial Strains
- Plasmid extraction
We are artificially extracting plasmids.
April 15 – April 29
- Still extracting plasmids
Do a great job, workstation—we know you can do it!
April 30 – May 12
- Yet again, we're extracting plasmids.
We're currently centrifuging, and we'll service our workstation.
May 12 –June 12
Functional testing of bacteria And Plasmid resource mining
- During this period, we will conduct stress culture tests,including low temperature, high temperature, acid, alkali, and salt,on the natural plasmid-containing bacteria that were screened between March 31 and May 12.
- Continue the plasmid resource mining work
The bacteria under "inspection" are currently in shake flasks.
June 12 – July 12
Functional testing of bacteria
- During this period, we cultured natural bacteria containing plasmids under stress conditions, including low temperature, high temperature, acid, alkali, and salt.
We are currently inoculating bacteria into the culture medium
June 12 – August 12
Robustness testing of recombinant engineered bacteria
- Natural plasmids were introduced into the E. coli BL21 strain to construct recombinant engineered bacteria, which were then subjected to stress test cultivation. The stress conditions included low temperature, high temperature, acid, alkali, and salt respectively.
June 16 – July 16
ORF Prediction and Functional Verification
- ORF Prediction and Functional Verification.Functional verification included stress cultivation tests and SDS-PAGE electrophoresis analysis.
We are currently using a PCR instrument to construct recombinant engineering bacteria.
July 17 – August 17
- Yet again，ORF Prediction and Functional Verification.
We are currently performing heat shock transformation.
August 18 – September 18
- It's finally over! ORF Prediction and Functional Verification.
April 5 – April 12
Background research
- This week we focused on some of the paperwork and the groundwork for the experiments ahead
July 7 - July 14
Comparative analysis of bacterial species
- Classify bacterial strains according to sampling locations
- Classify bacterial strains according to genus-species relationships
Sequence alignment analysis
- Complete sequence determination of plasmids
- Sequence alignment and analysis of plasmids
June 9 - August 11
- ORF predictionDetermine the ORF sequences
- Conduct analysis and comparison of the ORF sequences
We feel a sense of joy looking at the map we built.
A smooth and efficient spotting (Ｔ▽Ｔ)
Found a misplaced pipette tip Σ(ﾟ∀ﾟﾉ)ﾉ
I prepped a whole bunch of culture medium, but it ended up getting contaminated—guess I need to step up my game with sealing and sterilization checks!(╥_╥)
I washed a deadly amount of bottles again today—seriously, why do there have to be so many?!（╯－＿－）╯╧╧
Extracting plasmids is just so tedious—seriously, it’s draining.╮(︶﹏︶)╭
Every step of this experiment was steeped in the accumulation of "untiring efforts": In order to catch up with the schedule, team members lingered around the workbench for several consecutive days. The unwashed lab bottles piled up like a small mountain, and the prepared culture medium would be rendered useless by accidental bacterial contamination in the blink of an eye. Yet the results always fell a notch short of expectations, even the pen used to record data seemed to carry a touch of heaviness.
On several occasions, when faced with piles of samples and unsatisfactory graphs, everyone had the thought of "should we pause for a while?" But no one actually stopped — some volunteered to work overtime to optimize the experimental process, while others pored over numerous literatures to adjust the extraction parameters. Even if each improvement was only a tiny step, they persisted in gathering these trivial efforts bit by bit.
It was precisely this little bit of perseverance and repeated refusal to give up that, like the footprints left by caravans step by step along the ancient Silk Road, turned the initial scattered attempts into a coherent operational framework. Later, this refined process extended to broader experimental directions. In the end, we not only achieved excellent results that exceeded expectations, but also gained a profound insight: There is no overnight success on the road of scientific research.
Those seemingly trivial accumulations and the perseverance in the face of difficulties are just like the perseverance of the Silk Road that crossed mountains and seas. They will eventually connect "dots" into "lines" and expand "lines" into "planes", fostering true value through breakthroughs. This growth has become the most precious asset on our scientific research journey.