@@jacksonlaboratory For this why don't we use strong detergents for cell lysis? and then enzymes for for degrading nuclear membrane so that DNA gets exposed?
@@ektagupta7504 STRONG detergents are used for cells with cell walls (and hard ones of course). This experiment's sample were saliva spat by students which is linked with animal cells. Animal cells do not have a cell wall only a cell membrane, hence strong detergent is not advised, scientifically to be used since it might quickly disrupt the cell membrane and denature the very genomic DNA we want to extract. Hence enzymes are mostly preferred...
Do you find it difficult to work with such loose fitting gloves? Trying to work fast and one handed maneuvers causes your gloves to get caught inside the eppendorf tubes when you close them.
Hello Thenk you so much for the crystal clear method demonstration. I have question regarding this. We can crushing in nitrogen then how much time we store these material??? Like one month or 15 days? Any idea about this.
Hey there... I am isolating plant dna from leaf sample and getting smear in it... Even after RNase treatment and purification of the DNA. Could you please provide suggestions regarding the removal the smear... Thank you in advance 🙏
All of these "proprietary" solutions involved with DNA are usually salt solutions. They make them proprietary to keep that a secret because people do not want to pay 50 bucks for an ounce of salt water.
Ignore my statement below; it is an incorrect conjecture that would need to occur later in the process and only if they were planning on performing gene splicing. They haven't explained what they plan to do after extracting the DNA. The sample is mixed with the purifying solution AFTER heating the sample. I believe that they are using it to break down and dissolve the cell walls. Then they begin the annealing process (the ice). From there they use the centrifuge to separate the lighter material (dna) from the heavier material (leftover dissolved larger scale cellular detritus - the white pellets). ---- Look up 'Genetic Engineering - Isolating a gene'. Now I am not an expert in this subject -- but if I had to guess; the 'purifying solution' contains either one or many types of 'Restriction Endonucleases' that are used to split the DNA into discrete pieces prior to putting the DNA into their gene sequencer.
Can I get the protocol DNA extraction for bacteria?please.
4 ปีที่แล้ว +1
Hi, write me to tomas.hluska at upol.cz I can translate our "home-made" method for you (I think it's from Sambrook anyway) that we used in our lab course.
Can somebody explain why sometimes we keep the supernatant and sometimes we throw it away. Like after you grow a culture and you want to store for -20C freeze, we centrifuge and discard the supernatant and keep the pellet. But in this case its the opposite.
In the case you're describing the pellet has the cells in it. In this case, the DNA is contained in the supernatant while impurities (protein, cell wall components, etc.) are contained in the pellet because the purifying solution added to the mixture (that made it cloudy) denatures the cells. The DNA will not precipitate out into a pellet unless you add specific reagents which usually come in another kit. Whether you keep the supernatant or not largely relies on your end goal, and what reagents and procedures you are using.
4 ปีที่แล้ว +2
That depends which part is important for you. In case of bacteria storage, you pellet the bacteria which you are interested in and thus proceed with the pellet. In case of DNA extraction, you actually use both (in two separate centrifugation steps; unless you use binding columns). First, you precipitate and sediment the proteins which you want to get rid of, while your DNA is in the solution. Hence you work with the supernatant. In the second step, you precipitate your DNA with alcohol and hence you proceed with the pellet.
If you are dispensing the pipette yes. It’s recommended to have it at a 45 degree angle and against the wall of the tube you are dispensing into. It will help “pull” all the volume out.
the detergent disrupts the cell and nuclear membranes of each cell to release the DNA. It does this by dissolving lipids and proteins that hold the membranes together.
The lysis step. SDS is what you would call the soap. This is one of the first steps, along with a enzyme that breaks down proteins. Then your isopropyl/etoh (alcohol), which precipitates out the DNA since it's not soluble in alcohol.
I know it is an old video. Anyone can tell me the purpose of purifying solution? could i use this protocol for E coli solution and for qPCR?
4 ปีที่แล้ว +1
E. coli has tough cell wall, so I don't think so. You need alkaline lysis for bacteria. qPCR requires clean DNA what should be provided by any kit method.
![Кто звонил ЗАХАРОВОЙ на самом деле? 😁 [Пародия]](/img/n.gif)
why do we do heat incubation in the beginning??
Thanks for the question. Heat incubation activates the enzyme that will break down the cells and nuclei to expose the DNA.
thanks :)
@@jacksonlaboratory For this why don't we use strong detergents for cell lysis? and then enzymes for for degrading nuclear membrane so that DNA gets exposed?
I think to sterilize it good question
@@ektagupta7504 STRONG detergents are used for cells with cell walls (and hard ones of course). This experiment's sample were saliva spat by students which is linked with animal cells. Animal cells do not have a cell wall only a cell membrane, hence strong detergent is not advised, scientifically to be used since it might quickly disrupt the cell membrane and denature the very genomic DNA we want to extract. Hence enzymes are mostly preferred...
If you’re in the 23andMe batches that arrived between 29 November and 9 December, this simply process takes seven weeks (and counting).
It is a very helpful video for the beginners .....................who want to start research on DNA analysis
nah fam, you vortex 2 or 3 at the time if you gangsta
Anybody here after Biotech experience
Lol, yes I am 😂
Running student
this video is easy to procedure level learnig help to experimental research
It would be very good practice if tubes were labelled from the very start.
Good practice!!
That disposal cup could be a source for contamination of the sample.
Can anyone agree that pipetting is preatty satisfiying?
Hi, can you please make such a wonderful video for DNA Extraction from soil samples?
Comment by Allen student entusepro 1st batch saurav Kumar yadav .jai hind
unfortunatly it does not talk about the kit in the begining of the video
Do you find it difficult to work with such loose fitting gloves? Trying to work fast and one handed maneuvers causes your gloves to get caught inside the eppendorf tubes when you close them.
How many available methods are there for DNA extraction from teeth?
thank you
it's so useful.Thanks
what is the purpose of 90 min incubation?
Maa shaa Allah, Thank you :)
Are there any special ingredients or protocol steps required for DNA extraction for RADseq projects?
Hello
Thenk you so much for the crystal clear method demonstration.
I have question regarding this. We can crushing in nitrogen then how much time we store these material??? Like one month or 15 days?
Any idea about this.
Hi does the buffer break down the lipids in the cell membrane as does using detergent?
Hey there...
I am isolating plant dna from leaf sample and getting smear in it...
Even after RNase treatment and purification of the DNA.
Could you please provide suggestions regarding the removal the smear...
Thank you in advance 🙏
thank you for video
What is the Kit used here?
hello, what is the solution that the saliva goes into while inside the falcon tube?
Vanakam da mapla
thank you this is so helpful
Why do we incubate on ice??
What does the purifying solution used consist of ?
All of these "proprietary" solutions involved with DNA are usually salt solutions. They make them proprietary to keep that a secret because people do not want to pay 50 bucks for an ounce of salt water.
I am coming from DNA typing lab classroom
What method was used in this DNA extraction?
For accuracy
good video
Thanks for sharing this video
Thank you so much DR RORPOPOR HERBAL on TH-cam you saved my life from this deadly PCR virus, I got cured within 14daysly herpesly herpes..........
Which experiment u perform?
is it same protocol for extraction of Theileria annulata DNA extraction from animal blood sample?
ITS VERRY IMPORTANT TECHNOLOGY
What are the chemicals used for DNA extraction?
do we follow the same protocol when extracting DNA from human hair?
Same question
It all depends on the protocol different extraction kits have slightly different protocol but most are similar
Bonjour, si possible je veux Protocole d'extraction de l'ADN a partire de bacterie
what materials where used?
How extract DNA whataman filter paper
Wow, my lab looks like a hobo dumpster compared to this...
What is the purifying solution?
Hi Trasvi. The purifying solution is a proprietary solution from DNAGenotek that precipitates cell debris as part of their saliva extraction kit.
Ignore my statement below; it is an incorrect conjecture that would need to occur later in the process and only if they were planning on performing gene splicing. They haven't explained what they plan to do after extracting the DNA. The sample is mixed with the purifying solution AFTER heating the sample. I believe that they are using it to break down and dissolve the cell walls. Then they begin the annealing process (the ice). From there they use the centrifuge to separate the lighter material (dna) from the heavier material (leftover dissolved larger scale cellular detritus - the white pellets).
----
Look up 'Genetic Engineering - Isolating a gene'. Now I am not an expert in this subject -- but if I had to guess; the 'purifying solution' contains either one or many types of 'Restriction Endonucleases' that are used to split the DNA into discrete pieces prior to putting the DNA into their gene sequencer.
Salt water
just procedure ?
From what samples was the DNA extracted from?
I would rather use a Kit such as qiagen or zymo
Why did u put it in ice
Can I get the protocol DNA extraction for bacteria?please.
Hi, write me to tomas.hluska at upol.cz I can translate our "home-made" method for you (I think it's from Sambrook anyway) that we used in our lab course.
This video was almost perfect but they had to choose that background music.
Anak Fk unipa semangat 🔥
Can somebody explain why sometimes we keep the supernatant and sometimes we throw it away. Like after you grow a culture and you want to store for -20C freeze, we centrifuge and discard the supernatant and keep the pellet. But in this case its the opposite.
In the case you're describing the pellet has the cells in it. In this case, the DNA is contained in the supernatant while impurities (protein, cell wall components, etc.) are contained in the pellet because the purifying solution added to the mixture (that made it cloudy) denatures the cells. The DNA will not precipitate out into a pellet unless you add specific reagents which usually come in another kit. Whether you keep the supernatant or not largely relies on your end goal, and what reagents and procedures you are using.
That depends which part is important for you. In case of bacteria storage, you pellet the bacteria which you are interested in and thus proceed with the pellet.
In case of DNA extraction, you actually use both (in two separate centrifugation steps; unless you use binding columns). First, you precipitate and sediment the proteins which you want to get rid of, while your DNA is in the solution. Hence you work with the supernatant. In the second step, you precipitate your DNA with alcohol and hence you proceed with the pellet.
Are you supposed to pipette
at a slanted angle like that?
If you are dispensing the pipette yes. It’s recommended to have it at a 45 degree angle and against the wall of the tube you are dispensing into. It will help “pull” all the volume out.
I was looking for NaCl Ctab extraction but i think this is the boiling method
What are the thing needed for extract wher we can buy please reply
Ok
GOOD METHODS
How do you remove RNA from DNA? Won’t RNase simply break it down into nucleotides?
Why we put the solution in ice?
Can i extract DNA from cooked meat?
Short answer - yes! www.ncbi.nlm.nih.gov/pubmed/18791146
English is so difficult can not got fruitful knowledge.🥺🥺
when will protocol 2 come???
Rabin Subba
Angur Ali 👧👧😉🚧↕🚩🚩🚩🚩⚠🚩🚩🚩⚠🚩⚠🗿🗿🏣xcxch^^*^&&&&&;&&
Angur Ali -iklllĺloııkı^oıooooooooooooooooöolöıl.,ll i
The Jackson Laboratory çllşşş
th-cam.com/video/QYpX94prb0A/w-d-xo.html Protocol 2 - PCR
please what organism dna extraction is this?
Human saliva
Hi! what kind of DNA is that? (genomic, plasmidial...)
It is DNA from mouse tissue.
@@jacksonlaboratory from mouse tissue? Didn't you say that students spit into the tubes?
@
The students did spit in the tubes, but they were instructed to chew the mice first.
Hi what do extraction DNA vibrio cholera manul?
My DNA beku future,DNA to life future.kotak es beku
To life future.
Tofan Andriansah AE
Sorry but the background music is distracting not to mention annoying as hell. I could not finish watching it.
You aren't kidding--a country song for a high-tech topic.
Masa 3.000 masehi ada.
We extract dna form hair
For dna extraction from hair, is it the same method as this?
Why are the samples put in ice for ten minutes before centrifuging out the impurities?
For accuracy
cold reduces solubility of impurities.
Hi!
We did DNA extraction from kiwi fruit and we used hand soap with the mixture. My question is which function has the soap in the lab?
the detergent disrupts the cell and nuclear membranes of each cell to release the DNA. It does this by dissolving lipids and proteins that hold the membranes together.
leyna yusof thx ❤️
The lysis step. SDS is what you would call the soap. This is one of the first steps, along with a enzyme that breaks down proteins. Then your isopropyl/etoh (alcohol), which precipitates out the DNA since it's not soluble in alcohol.
Can this method be used in maize?
I don't think so. Plants have cell wall, which need to be broken. Often various protocols using CTAB are used.
most likely.
the pipette should not be on the bench
I know it is an old video. Anyone can tell me the purpose of purifying solution? could i use this protocol for E coli solution and for qPCR?
E. coli has tough cell wall, so I don't think so. You need alkaline lysis for bacteria.
qPCR requires clean DNA what should be provided by any kit method.
@ or heat... Bacteria break apart easily, 10 min at 95C
@@AndrewLyon23 we used heat for direct genotyping bacteria by PCR, but does qPCR work as well?
Labeling of tubes was not done when placing in ice need to redo video
high tech potions class
This video is very usually found me
Good times
whats up wer u at velez pipzz
Is it possible to extract DNA from dead people saliva
why do you want to extract DNA from dead?
A little bit too suspicious buddy
@@avolatube_8253 ... forensics
are you asking for a friend?
song? lol
فد فتهمنا😑😑😑😑
당신이 판촉하는 항목을 구입하는 방법
fal
Zero breast cancer awareness
Anlamadım
If blood observed on nasal swab kit during taking sample from nasal, it will be cause of covid-19
ga asik part-part an