@@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.
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 🙏
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.
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.
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 года назад+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.
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 года назад+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.
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.
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 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 года назад+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.
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...
It is a very helpful video for the beginners .....................who want to start research on DNA analysis
If you’re in the 23andMe batches that arrived between 29 November and 9 December, this simply process takes seven weeks (and counting).
Can anyone agree that pipetting is preatty satisfiying?
It would be very good practice if tubes were labelled from the very start.
Good practice!!
Hi, can you please make such a wonderful video for DNA Extraction from soil samples?
nah fam, you vortex 2 or 3 at the time if you gangsta
this video is easy to procedure level learnig help to experimental research
That disposal cup could be a source for contamination of the sample.
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.
Anybody here after Biotech experience
Lol, yes I am 😂
Running student
Yes
it's so useful.Thanks
How many available methods are there for DNA extraction from teeth?
unfortunatly it does not talk about the kit in the begining of the video
Comment by Allen student entusepro 1st batch saurav Kumar yadav .jai hind
Are there any special ingredients or protocol steps required for DNA extraction for RADseq projects?
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 🙏
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.
is it same protocol for extraction of Theileria annulata DNA extraction from animal blood sample?
I am coming from DNA typing lab classroom
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
what is the purpose of 90 min incubation?
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.
What are the chemicals used for DNA extraction?
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.
thank you this is so helpful
Maa shaa Allah, Thank you :)
hello, what is the solution that the saliva goes into while inside the falcon tube?
What method was used in this DNA extraction?
For accuracy
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
What is the Kit used here?
Why do we incubate on ice??
How do you remove RNA from DNA? Won’t RNase simply break it down into nucleotides?
Thanks for sharing this video
Thank you so much DR RORPOPOR HERBAL on RUclips you saved my life from this deadly PCR virus, I got cured within 14daysly herpesly herpes..........
From what samples was the DNA extracted from?
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.
Bonjour, si possible je veux Protocole d'extraction de l'ADN a partire de bacterie
How extract DNA whataman filter paper
I was looking for NaCl Ctab extraction but i think this is the boiling method
Which experiment u perform?
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.
thank you for video
I would rather use a Kit such as qiagen or zymo
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.
Can i extract DNA from cooked meat?
Short answer - yes! www.ncbi.nlm.nih.gov/pubmed/18791146
ITS VERRY IMPORTANT TECHNOLOGY
when will protocol 2 come???
Rabin Subba
Angur Ali 👧👧😉🚧↕🚩🚩🚩🚩⚠🚩🚩🚩⚠🚩⚠🗿🗿🏣xcxch^^*^&&&&&;&&
Angur Ali -iklllĺloııkı^oıooooooooooooooooöolöıl.,ll i
The Jackson Laboratory çllşşş
ruclips.net/video/QYpX94prb0A/видео.html Protocol 2 - PCR
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.
Why we put the solution in ice?
what materials where used?
good video
What are the thing needed for extract wher we can buy please reply
Ok
My DNA beku future,DNA to life future.kotak es beku
To life future.
Tofan Andriansah AE
just procedure ?
Why are the samples put in ice for ten minutes before centrifuging out the impurities?
For accuracy
cold reduces solubility of impurities.
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?
Hi what do extraction DNA vibrio cholera manul?
thank you
GOOD METHODS
Why did u put it in ice
Wow, my lab looks like a hobo dumpster compared to this...
We extract dna form hair
For dna extraction from hair, is it the same method as this?
please what organism dna extraction is this?
Human saliva
English is so difficult can not got fruitful knowledge.🥺🥺
Anak Fk unipa semangat 🔥
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.
Vanakam da mapla
This video was almost perfect but they had to choose that background music.
the pipette should not be on the bench
Masa 3.000 masehi ada.
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?
Labeling of tubes was not done when placing in ice need to redo video
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.
This video is very usually found me
high tech potions class
whats up wer u at velez pipzz
فد فتهمنا😑😑😑😑
당신이 판촉하는 항목을 구입하는 방법
song? lol
Good times
Anlamadım
If blood observed on nasal swab kit during taking sample from nasal, it will be cause of covid-19
Zero breast cancer awareness
ga asik part-part an
fal