RNA Extraction | acid guanidium thiocyanate phenol chloroform extraction

[@charleszhou5021]

We are supplier of guanidine thiocyanate. it is a very useful vedio,Thanks!

[@aisasam8303]

watching your videos is better than my lectures . you’re actually teaching and you sound like you’re about it! thank you

[@shomusbiologyofficial]

You're welcome

[@anilkumarraushan6483]

Ya

[@monicatorres3366]

I follow your videos becuase for the most part they are great, very informative. However you make a big mistake in this video by stating that phenol separates from chloroform. What really happens is that the solution is separated into two phases: the aquous phase with water and RNA (under acidic conditions) and the phenol/chloroform phase which dissolves proteins and DNA (phenol interphase) and lipids (chloroform)

[@Shahid330050]

Thanks Monica for clarification otherwise I was also confused to know that phenol is in aqueous phase.

[@tultu1]

Of course Phenol remains separated from chloroform, but he should say phenolic phase instead of aqueous phase. It removes confusion.

[@charleszhou5021]

Great ,very professional, hope we can discuss further, thanks!

[@PraveenKumar-co2vl]

Deepinder Sokhi, you are right! CHCl3 & PhOH both are organic solvents, but small variations, phenol is soluble in water to some extent. It is due to its ability to form hydrogen bonding with water molecules. However the large part of phenol molecule is phenyl group that is non polar and hence its solubility if limited in water. to Hydrogen bonding nature of phenols (-OH). The cell components are two types polar and non polar. Polar components go in to phenol layer (top) and non polar components go in to CHCl3 layer (bottom). Polar components: majorly RNA/DNA (Note: Having 2'OH gp in RNA makes more polar than DNAs, DNA resolves at the interface) Non polar components: Proteins (depends on "R" groups) & Lipids

[@linamahmoud651]

can you give me a link that explains this briefly ?

[@babitabhatt6469]

Well explained ...Thank u sir😊..Are u that praveen kumar ... Author of pathfinder?🤔🤔🤔🤔🤔

[@nidapraveen3759]

Only youtuber thay makes videos on topics like this that no one can..

[@gregorysmith1134]

Beautifully explained. Thanks for sharing.

[@shomusbiologyofficial]

You're welcome

[@anupamagoswami]

Thank you shomu, tu god hai bhai! love from IIT-M.

[@charleszhou5021]

We are producer of guanidine salts in China.Guanidinium thiocyanate is one of our strong products.hope we can discuss further.thanks!

[@shomusbiologyofficial]

You're welcome

[@charleszhou5021]

@@shomusbiologyofficial What is your business!

[@anilkumarraushan6483]

@@charleszhou5021 teaching

[@fahadshah8057]

I have no words to say thank .you solve my big problem.

[@Hj-qm1uy]

thank you so much I couldnt figure out the reason why rna and dna cann be seperated

[@shomusbiologyofficial]

You're welcome

[@swetchamangalagiri2279]

Hello sir, can u explain DNA extraction method from meat sample

[@nimbappar9983]

U r genius sir .very brilliant teaching.thank u sir very much

[@giahungtran6822]

Thank you so much for a very informative and concise lecture!

[@shomusbiologyofficial]

Thank you

[@jyotijadhav2472]

Excellent sir

[@shomusbiologyofficial]

Thank you

[@zaidabdullah8316]

You are awesome man , 🌺 from iraq

[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures

[@syedishu2807]

Shomu sir i need your help,i am working in the pcr lab from last few days my Rna extraction gets repeated ,i reextract the Rna what could be the possible factors that interfere in my RNA extraction

[@garimaanand9680]

You are amazing

[@manorikuruppu8989]

Your effort is much appreciated. Thanks so much for your informative details. Could you please let me know how to extract high quality RNA for metagenomics

[@muhammetnazar1754]

woow legend. Thanks.

[@shomusbiologyofficial]

You're welcome

[@dylanyip-chuck8861]

Thanks. You are a very good teacher.

[@shomusbiologyofficial]

+Dylan Yip-Chuck thank yiub

[@rameshram-hm3jh]

Well said sir keep doing

[@shomusbiologyofficial]

Thank you

[@sagartyagi1229]

Sir... isn't phenol organic? And if it is organic then how come it represent aqueous phase in phenol:chloroform gradient? Referring to 7:10 min in video.

[@saucycreates1076]

I wanted to know the purpose of TriZol ? Does it give Acidic environment

[@emadbendahman3438]

Thanks so much. Is there a lecture in steps of DNA extraction?

[@subhamsaha2235]

top class....nice man

[@shomusbiologyofficial]

+Subham Saha thank you.

[@lesquisse8898]

Thanks a lot!

[@shomusbiologyofficial]

You're welcome

[@adyashasarangi2002]

Please help me out.... If dna rna and protein are placed in testtubes...then how to distinguish them without disturbing them?

[@najatabdulrahim1711]

I want to ask for the reason why in the acidic conditions RNA will be in the aqueous phase while DNA will be in the organic phase ?

[@lachhamapradhan4218]

Is this process same for both animal and plant tissue?

[@cholponzhakshylykova4099]

thanks, nice explanation :)

[@shomusbiologyofficial]

You're welcome

[@komals.choudhari9818]

Sir graph kaise represent karna hai ye batao na

[@muhammadali-ly1kp]

Great

[@ProscoviaKanyunyuzi]

Why does the RNA and DNA separate in aqueous phase

[@AKSHAYDESHMUKH-me6zx]

Thank you sir

[@shomusbiologyofficial]

You're welcome

[@sumeshsudhakaran4823]

How will you separate prokaryotic mRNA? Suppose if we have to isolate rRNA and tRNA what will we do? One doubt too...when we isolate DNA it might contain mitochondrial and chloroplast DNA with genomic DNA right? How will we eliminate those DNA?

[@ruchikachoraria7837]

Hello Sir, You are a great teacher and have helped us so much! thank you .. but here you are mentioning that phenol is aqueous?? both chloroform and phenol are organic solvent btw

[@ahasanulshah249]

If you don't mind... Subrata das describe something different about aqueous inorganic portion and organic portion quite differently.. I can't understand properly..

[@vaishnavibhardwaj9970]

still don't understand the experiment i mean i understand each of every step but don't understand which step 'll come first can u explain me here sirr plzzz also wanna know that this process is also called M_rna enrichment???

[@vanessasmith4637]

Thanks man.

[@syedahsanshah8344]

Grwat teacher

[@shomusbiologyofficial]

You're welcome

[@soliangmanyu2864]

Sir what would be the position of DNA&RNA ,if the Ph is basic?

[@radhekrishnamotivationgang6115]

sir why at acidic ph only dna comes in organic phase?

[@ErikVenosa]

nice job

[@priyanshimishra6667]

what will happen if the pH is basic?

[@royjoseph1498]

best ever!!!

[@ridoy1992]

awesome

[@sabahatmajid9599]

Great explanation. One question: how do we keep the pH acidic?

[@MalikAli-ck8sq]

why acidic pH causes RNA to stay in aqueous phase while forces DNA in organic phase ?

[@zaidabdullah8316]

Malik Ali cos I think Rna has sugar molecule that lack hydrogen while dna has not lack hydrogen molecule. Mean dna more acidic

[@sagartyagi1229]

Dear Malik...it has been 4 years since you asked this question and I hope you had got the answer of it and if not here it is....... First of all Both RNA and DNA are Polar in nature because of negatively charged phosphate backbone which in the presence of acidic conditions gets neutralized. Now, since RNA has exposed bases unlike DNA where because of double helix they are present inwardly, RNA molecules remain positively charged because of its bases and this makes RNA soluble in aqueous phase and DNA being neutral gets no place in aqueous as well as Organic phase hence it comes out to be in middle.

[@sudiptajana1854]

@@sagartyagi1229 how rna is positively charged in acidic conditions..can you elaborate this line what you have written?

[@sagartyagi1229]

@@sudiptajana1854 Hye....because of the protonation (under acidic condition) of nitrogen groups which are integral part of nitrogenous bases.

[@sudiptajana1854]

@@sagartyagi1229 ook..got it..and the phosphate groups are protonated in both the DNA and RNA...ri8?or this is only for the DNA?

[@sulecanberk7608]

perfect

[@deepindersokhi7441]

phenol is not inorganic that's an organic compound.....

[@bikidas6690]

How guanidium breaks rnase??

[@ranabasit4903]

how can be aquous phase separated? is it with help of pippet or something else?

[@JP-wx6uh]

Spinning it down in a centrifuge

[@ArnavPadhi]

How to isolate total mRNA from a pool of RNA?

[@shiprasiva9495]

You can use the column purification method where oligo dT resins are used to separate mRNA from other RNAs. Because of the poly A chain end at 3'

[@JP-wx6uh]

Why would Chloroform settle on the bottom? It doesn't weigh nearly as much as Phenol. Phenol's molecular weight is like 400-something atomic mass units versus Chloroform which is like 3 times less.

[@shantanusingh8267]

It's all about density, as the density of chloroform is 1.49g/cm cube white that of phenol is 1.07g/cm cube

[@vaalwyn]

what happens if i isolate proteins from a tissue sample using this method? will i be able to quantify the proteins by ELISA? If the proteins get denatured during extraction, and can these proteins be detected by ELISA? i know it has nothing to do with RNA extraction but if you can help ill gladly appreciate it. :)

[@Randhirkumar-hl5ux]

hindi verjan

[@mimimom7666]

ph buffer

[@yayirce5032]

Cortese el pelo gei

[@indoremedicine8424]

Cloroform kisko chahiye

[@Geambasu169]

I dont understand your english. Maybe this course it is for your indian friends. Try to learn how to be more universal and talk for everybody. To do that learn (even you) how to speak without freakin accent. Keep up the good work.

[@anshulrana6211]

Thank you sir

[@shomusbiologyofficial]

You're welcome

[@chanellekuete3503]

Thanks you❤

[@shomusbiologyofficial]

You're welcome