RNA Extraction and Purification |Acid Guanidinium Thiocyanate Phenol Chloroform (TRIzol) extraction|

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Acid guanidinium thiocyanate-phenol-chloroform extraction (abbreviated AGPC) is a liquid-liquid extraction technique used to extract RNA molecule .
Procedure:
1)cell lysis by GTC buffer ( Guanidinium thiocyanate )
2)sodium acetate buffer (pH=4)
3)phenol chloroform extraction
4)isopropanol precipitation
5)70% ethanol wash
6)store RNA
This method relies on phase separation by centrifugation of a mixture of the aqueous sample and a solution containing water-saturated phenol and chloroform, resulting in an upper aqueous phase and a lower organic phase (mainly phenol). Guanidinium thiocyanate, a chaotropic agent, is added to the organic phase to aid in the denaturation of proteins (such as those that strongly bind nucleic acids or those that degrade RNA). The nucleic acids (RNA and/or DNA) partition into the aqueous phase, while protein partitions into the organic phase. The pH of the mixture determines which nucleic acids get purified.[3]Under acidic conditions (pH 4-6), DNA partitions into the organic phase while RNA remains in the aqueous phase. Under neutral conditions (pH 7-8), both DNA and RNA partition into the aqueous phase. In a last step, the nucleic acids are recovered from the aqueous phase by precipitation with 2-propanol or ethanol.
Guanidinium thiocyanate denatures proteins, including RNases, and separates rRNA from ribosomal proteins, while phenol, isopropanol and water are solvents with poor solubility. In the presence of chloroform or BCP (bromochloropropane), these solvents separate entirely into two phases that are recognized by their color: a clear, upper aqueous phase (containing the nucleic acids) and a lower phase (containing the proteins dissolved in phenol and the lipids dissolved in chloroform). Other denaturing chemicals such as 2-mercaptoethanol and sarcosine may also be used.

[@mahibarot2828]

Great detailed explanation! Thank you!

[@rajinderkumar36272]

Very useful and understandable thank u so much 🎉

[@namratasamanta452]

Khub upikrito holam . Sir khub sundor bojhan aapni

[@downtoearthveeresh2790]

Point to point explain. Easy to learn

[@ShivaniSharma-ni6nb]

U r amazing sir u hv explained each and everything so nicely thank u so much keep making such videos that will help all students

[@misbahfarooq3747]

Thank you!

[@jahnavip1307]

Helped alot during exams...👍

[@XianOrtiz7]

great job

[@toysspirits995]

Amazing explantion😍

[@13violalover]

Very well explained--thank you for making this video!

[@harshikasharma3812]

Very nice video..this is very helpful to me..nice explanation...😍😍

[@kapilraghuwanshi1092]

It's really helpful.

[@sanashaikh984]

Very well explained
Thanks

[@suhitashow6789]

Thank you dada.this is most helpful video.

[@bhargaviparmar4411]

very nice.,..., you explain why ? , what ? when ? so understood basics.., thank you sir

[@microbioscope18]

Bhargavi Parmar .... Most welcome

[@charleszhou5021]

Very good video, we are producer of guanidine thiocyanate.hope we can discuss further, thanks!

[@satishkoyidala5538]

Very useful to us thank you bro

[@pratikshachandorkar6754]

Thank u so much sir... Very helpful video... Nicely explained..

[@paulnisa875]

Fantastic explanation

[@debranichakraborty6004]

this video saved me.

[@anaghav3906]

Thank you so much

[@mohammedwaseequrrahman6484]

Superb!

[@swarnalisarkar8216]

nice

[@sheezaandleeb8449]

well done

[@Kad991]

thank you

[@swarnalisarkar8216]

dunno why ur tutorials get so lesser amount of likes

[@user-bi7hl4lm4h]

great!!!!!!!

[@pranavbvn5537]

I have a doubt... wouldn't the organic layer be mostly chloroform? Or is it a mixture of Phenol and Chloroform?
Thanks in advance...

[@monikareddy3825]

Superb explanation...Thank u Sir🙏

[@melaniebillaud8404]

Juste génial !

[@bakhtawarbakhtawar799]

Good

[@nikhatlegendsquad1703]

Apke Vedio bohot ache hai laikin kuch word apke samjh Nahi ate kya Bolto Ho app... Srf kuch word

[@kapilraghuwanshi1092]

Please upload microarray technique

[@anamikagoyal2826]

Amazing sir but where will other intracellular impurities except dna nd protein will go . Will they go to organic phase along with dna ?

[@hibakaleed387]

Please I don't understand why rna is more acidic than dna can you help me??

[@mahibarot2828]

For this video purpose, RNA is acidic because it contain 2 OH group which are willing to donates its electron to the H+ proton of sodium acetates buffer. This will cause both oxygen present in pentose sugar to become positive hence cause overall RNA molecule to become acidic.

[@sandhiyakumaresan4589]

sir, thank you.... can you kindly provide the reference

[@abisulagam9045]

Please give the measurement also

[@razairam3412]

Kuch wording ki smjh ni a riii

[@abisulagam9045]

If you see the comment means please give measurement

[@ADULTFUN-uy9ip]

हिंदी में भी बताते तो aacha hota

[@cesarmadmax4170]

Shomus isomer XD

[@ramankumarlaldas8697]

Thank you!

[@resh1237]

Thank you!