Isolasi bakteri dengan teknik pengenceran dan penanaman secara spread plate
Summary
TLDRThis video tutorial demonstrates the process of bacterial isolation using serial dilution and spread plate techniques. It begins with preparing tools and materials, including urine samples, dilution tubes, and sterile equipment. The urine sample is serially diluted, mixed with a vortex mixer, and then plated onto agar media using a spreader. The spreader is sterilized with alcohol and flame before use. The final step involves incubating the plates at the appropriate temperature to grow bacterial colonies.
Takeaways
- π¬ The video demonstrates a method for bacterial isolation using serial dilution and spread plate techniques.
- π§ The sample used in the demonstration is urine.
- π§ͺ Preparation includes sterilized equipment and materials such as a Bunsen burner, alcohol sprayer, and a Vortex mixer.
- π₯ Aseptic technique is maintained by sterilizing the workspace with 70% alcohol and using a Bunsen burner flame.
- π¦ The process starts with taking 1 mL of urine sample using a micropipette.
- π‘οΈ The sample is then transferred to a dilution tube containing 9 mL of sterile water, creating a 10-fold dilution.
- π The dilution tube is homogenized using a Vortex mixer to ensure even distribution of the sample.
- π This dilution process is repeated through a series of tubes to achieve higher dilutions.
- π The final dilution is plated onto an agar medium using a spreader, previously sterilized with 70% alcohol and a Bunsen burner.
- π‘οΈ The spreader is allowed to cool before use and is sterilized again after use.
- π± The petri dishes are incubated at the appropriate temperature to allow for bacterial growth.
Q & A
What is the main topic of the video?
-The main topic of the video is the isolation of bacteria using a technique called serial dilution and the spread plate method.
What is the first sample used in the video?
-The first sample used in the video is urine.
What are the materials needed for the serial dilution process as mentioned in the video?
-The materials needed include sterile water, a pipette, a Vortex mixer, a Bunsen burner, alcohol spray, and a disposable loop.
What is the purpose of using a Bunsen burner in the process?
-The Bunsen burner is used to flame the disposable loop and other tools to sterilize them and maintain aseptic conditions during the process.
How much urine sample is taken initially for the process?
-One milliliter (1 mL) of urine sample is taken initially using a micropipette.
What is the purpose of adding the urine sample to the first dilution tube containing sterile water?
-The urine sample is added to the first dilution tube to create a 1:10 dilution, which helps in isolating and counting the bacteria present in the sample.
How is the mixture homogenized after adding the urine sample to the dilution tube?
-The mixture is homogenized using a Vortex mixer to ensure the sample is evenly distributed in the dilution medium.
What is the next step after homogenization in the serial dilution process?
-The next step is to take 1 mL of the homogenized mixture and transfer it to the next dilution tube containing 9 mL of sterile water, and then repeat the homogenization process.
How many dilution tubes are used in the process, and what is the final step after the last dilution?
-Four dilution tubes are used in the process. After the last dilution, the final step is to perform the spread plate method by taking 0.1 mL of the last dilution and spreading it onto an agar plate.
What is the purpose of sterilizing the spreader before spreading the sample on the agar plate?
-The spreader is sterilized by flaming it over a Bunsen burner to prevent contamination of the agar plate with unwanted microorganisms.
What is the final step after spreading the sample on the agar plate?
-The final step is to incubate the petri dishes containing the spread plates at the appropriate temperature to allow the bacteria to grow.
Outlines
π¬ Bacterial Isolation Process
This paragraph describes a method for bacterial isolation using serial dilution and spread plate techniques. The process begins with preparing urine samples and various equipment including dilution tubes, sterile water, pipettes, a Bunsen burner, a Vortex mixer, and an alcohol sprayer. The workspace is sanitized with 70% alcohol, and a Bunsen burner is used to maintain aseptic conditions. Urine samples are taken and diluted in a series of dilution tubes, each containing sterile water, and mixed using a Vortex mixer. The dilution process is repeated until the fourth dilution tube. The final step involves spreading 0.1 mL of the last dilution onto an agar plate for incubation.
π‘οΈ Spread Plate Technique and Incubation
This paragraph details the spread plate technique for evenly distributing the bacterial sample on an agar medium. A sterile spreader, or drugalsky, is used after being cooled and sterilized with 70% alcohol and heated over a Bunsen burner. The spreader is used to evenly distribute the 0.1 mL of bacterial solution on the agar plate. After use, the spreader is again heated to sterilize it before storage. The final step is to cover the petri dish and incubate it at the appropriate temperature to allow bacterial growth. The video concludes with a thank you note, hoping the information is useful.
Mindmap
Keywords
π‘Isolation
π‘Serial Dilution
π‘Spread Plate
π‘Urine Sample
π‘Sterile Aquadest
π‘Micropipette
π‘Vortex Mixer
π‘Bunsen Burner
π‘Agar Medium
π‘Inoculation
π‘Incubation
Highlights
Introduction to bacterial isolation using gradient dilution and spread plate techniques.
Preparation of tools and materials including urine sample, dilution tubes, and agar media.
Use of Google Sky for media preparation.
Sterilization of equipment and workspace with 70% alcohol.
Ignition of Bunsen burner for aseptic work.
Preparation of urine sample using a micropipette.
Transfer of urine sample to the first dilution tube containing sterile water.
Homogenization of the sample using a Vortex mixer.
Sequential dilution process using new pipettes for each step.
Transfer of diluted sample to subsequent dilution tubes.
Final dilution step before plating.
Use of spread plate method for sample plating.
Sterilization of the spreader (drugalsky) with 70% alcohol and Bunsen burner.
Spreading of 0.1 mL of the final dilution onto the agar medium.
Even distribution of the sample on the medium using a cooled spreader.
Re-sterilization and storage of the spreader after use.
Incubation of the petri dishes at the appropriate temperature.
Conclusion and expression of gratitude for the tutorial.
Transcripts
Halo
assalamualaikum warahmatullahi
wabarakatuh pada video ini dijelaskan
cara isolasi bakteri menggunakan teknik
pengenceran bertingkat dan penanaman
sampel dengan metode spread plate sampel
yang digunakan yaitu urine pertama-tama
kita persiapkan alat dan bahan yang
pertama sampel urine kemudian tabung
pengenceran media agar Google Sky
jarum Ose
mikropipet ukuran seribu dan seratus
mikroliter
bluti
[Musik]
bunsen korek api murah pink
tisu
alkohol sprayer
dengan Vortex mixer dan tempat sampah
cara kerja dimulai dengan misteri Khan
area meja kerja dengan alkohol 70%
kemudian area meja kerja di Lab
menggunakan tisu
selanjutnya nyalakan api bunsen untuk
membantu kerja aseptis
kemudian persiapkan alat-alat yang akan
digunakan
[Musik]
langkah selanjutnya ambil sampel urine
sebanyak satu ML menggunakan mikropipet
[Musik]
kemudian dipindahkan ke tabung
pengenceran yang pertama yang berisi
sembilan ML aquadest steril
kemudian dibuang blukid selanjutnya
tabung dihomogenkan menggunakan Vortex
mixer
selanjutnya masih dari tabung yang
pertama ambil larutan sebanyak satu ML
menggunakan tipe yang baru untuk
dipindahkan ke tabung pengenceran yang
kedua
[Musik]
kemudian dihomogenkan kembali lakukan
hal yang sama hingga tabung pengenceran
yang
[Musik]
Hai
senyum mu
[Musik]
Hai senyum
[Musik]
setelah selesai memindahkan larutan
hingga tabung keempat dilakukan
penanaman sampel menggunakan metode
scriptlet caranya ambil larutan sebanyak
0,1 ml dari tabung pengenceran yang
terakhir
[Musik]
kemudian
dipindahkan ke medium na lalu
diratakan menggunakan drugalsky
sebelumnya drugalsky dilakukan
sterilisasi terlebih dahulu dengan cara
menyemprotkan alkohol 70% kemudian
dipanaskan diatas api bunsen
Hai
[Musik]
berikut ini cara meratakan larutan yang
telah diteteskan di atas medium Ena
drugalsky didinginkan terlebih dahulu
sebelum gunakan untuk meratakan sampel
yang telah diteteskan di atas medium
enak
[Musik]
setelah selesai digunakan
Panaskan kembali drugalsky sebelum
disimpan kembali
Langkah terakhir yaitu Sil cawan petri
menggunakan uriping selanjutnya
dilakukan
inkubasi pada suhu yang sesuai
Hai Terima kasih semoga bermanfaat
[Musik]
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