RNA Extraction Tutorial

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  • Опубліковано 22 гру 2024

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  • @letsgo9433
    @letsgo9433 9 років тому +34

    Good video and explanation!
    FYI for those in comments, it's good to air-dry RNA by leaving the tube on its side (on a kimwipe), as this prevents contaminants from settling in the tube. Also good practice to make aliquots of your dH2O that you draw from, so you can limit the number of times you remove water directly from your DEPC water stock.

  • @arletteal7944
    @arletteal7944 8 років тому +46

    This is the most perfect thing I have ever watched

    • @BeeRich33
      @BeeRich33 4 роки тому

      Biochem is fun. Micro is funner.

  • @itsmejulia1
    @itsmejulia1 2 роки тому +1

    I am so thankful this video exists! You explained it really well!

  • @aakritipriya6670
    @aakritipriya6670 Рік тому

    Felt like I was a kid again learning through cute drawings and sketches ❤ really awesome 💖

  • @michaeltolkachov4749
    @michaeltolkachov4749 7 років тому +9

    One of the best tutorials a have seen so far! (and I saw a lot) Great job :)
    Cheers from the LMU Munich, I'll check the other videos ;)

    • @charleszhou5021
      @charleszhou5021 4 роки тому

      you are very professional.hope we can discuss further,my email:zgzhou@hebeismart.com

  • @MirandaXxRose
    @MirandaXxRose 4 роки тому +3

    This is such a great video. I love the animations and it is explained very well. Thank you.

  • @ssdiaries1602
    @ssdiaries1602 6 років тому +5

    Thank u so much for such a peaceful and informative video
    I just cleared all steps now by ur video..keep it up

  • @kokipays7969
    @kokipays7969 6 років тому +2

    This is awesome. i hope it works well for plant samples as well.

  • @alivia181
    @alivia181 Рік тому

    It is one of the best clearly explained with funny animation videos I have ever seen

  • @karenwang9974
    @karenwang9974 5 років тому +26

    the cutest rnase that ever liveddddd

  • @gautamkannan1909
    @gautamkannan1909 4 роки тому +2

    Nice video!!
    Can you please explain how pH less than 4 will allow only RNA to be in the aqueous layer and not DNA.
    thanks

  • @mochaticha5944
    @mochaticha5944 4 роки тому +3

    Clear and precise explanation! Thank you

  • @Yousha999
    @Yousha999 5 років тому +4

    Thank you so very very very much for this amazing and wonderful explanation 👍😎

  • @pichej22
    @pichej22 Рік тому

    Awesome video! I appreciate these types videos so much

  • @joannas610
    @joannas610 9 років тому +5

    This is wonderful. Thank you so much!

  • @jackypina5247
    @jackypina5247 2 роки тому

    Great video! I have a presentation on RNA isolation and this helped a lot!

  • @09prabhavikamble2
    @09prabhavikamble2 Рік тому

    Omg I understood every concept quite clearly thank you for making it it's so helpful 💜😍

  • @jackreidy6167
    @jackreidy6167 4 роки тому

    I'm a chemist and this made sense so good job and thanks!

  • @yijingwang7308
    @yijingwang7308 4 роки тому +1

    Many thanks for this informative video. But I have a question about the precipitation of isopropyl alcohol. I read from other papers that the function for isopropyl alcohol is to lower the dielectric constant so that RNA could precipitate. But not for neutralizing the ions of RNA. Could anyone help me with this question?

  • @charleszhou5021
    @charleszhou5021 4 роки тому

    Great!RNA Extraction may use guanidine thiocyanate is used for RNA isolation and can solubilize proteins

  • @girlschannel11
    @girlschannel11 6 років тому +1

    Hello,I did not understand what actually is the use of phenol in TRISOL.Its use is only to be an organic dissolver for the other polar molecules?Our professor in laboratory used to say this is to "hold" the debris and DNA.Also ,why and when do we put chloroform in the whole process?Also, EtOH has the same use of isopropanol?

    • @laurab1105
      @laurab1105 6 років тому +1

      Your professor is right by saying phenol-chloroform mixture holds down protein and hydrophobic lipids in the denser organic phase. . You add the chloroform (my lab uses BCP as it is less dangerous) straight after homogenising cells/tissue in trizol to separate the phases. Isoproponal is used to precipitate DNA/RNA as they are insoluble. To do this with EtOH you would need to add sodium acetate to perform a precipitation. Other times we use EtOH is to clean the pellet, not precipitate.

  • @JP-wx6uh
    @JP-wx6uh 5 років тому +1

    I thought the organic phase layer would be composed of mostly chloroform rather than phenol, because Chloroform is a much more dense and heavier compound than phenol. ??

  • @laetitiarialland9112
    @laetitiarialland9112 2 роки тому

    Thanks for this great theoretical ans practical explanation.

  • @TheMedicalBoss
    @TheMedicalBoss 9 років тому

    Very well explained, Much better than my tutor

  • @veekbassey1126
    @veekbassey1126 4 роки тому

    How do you prepare bacteria sample for RNA extraction if you have plated culture?

  • @_danfiz
    @_danfiz 8 років тому +1

    Very informative, but i have a question. After we put phenol chloroform and there's aqueous layer and organic layer, do we discard the organic layer first? And then add the isopropyl alcohol?

    • @ChangBenson
      @ChangBenson  8 років тому +6

      +nur hafizah The aqueous layer is the clear layer on top, the organic layer is the reddish layer on the bottom. Your RNA should be in the top aqueous layer and is simply transferred by pipetting into a new tube. The remaining organic layer in the old tube contains your unwanted cellular debris and DNA and can be discarded. The ethanol precipitation step of your desired RNA is then performed on the aqueous layer that you had transferred into the new tube.

  • @shikhi9455
    @shikhi9455 4 роки тому

    Many thanks for this informative video.please upload more informative video like this.

  • @shilpasharma4383
    @shilpasharma4383 6 років тому +1

    Hi...i m working on gene expression for that i hv to isolate plant RNA for cDNA preprtn...can u share protocol of this method...n plz tell me shud i go for this method or RNA extraction kit for plant RNA isolation...

  • @riyaadrahim3316
    @riyaadrahim3316 9 років тому +2

    Thanks this video is very informative!

  • @rehabalsaleh166
    @rehabalsaleh166 3 роки тому

    Absolutely informative! Thank you so so much!

  • @a.s9509
    @a.s9509 Рік тому

    you don't need to put the samples on ice and work on ice all the time?

  • @zamarawan8154
    @zamarawan8154 2 роки тому

    Can you please give the reference for the described method?

  • @berilkocak859
    @berilkocak859 7 років тому +1

    Thanks a lot for this video! It was helpful.

  • @gaurichaudhari1848
    @gaurichaudhari1848 3 роки тому

    which sample is more suitable rather than bacterial cell culture..please reply

  • @tusharsarve6586
    @tusharsarve6586 4 роки тому

    Can you please state the reason, why RNA is isolated at pH 4 and DNA at pH 5/6?

  • @reafdaw01
    @reafdaw01 9 років тому +1

    Thanks, that was very well explained.

  • @HimanshuSharma-oy9ss
    @HimanshuSharma-oy9ss 2 роки тому

    Nicely done and explained.

  • @drngabomichel8762
    @drngabomichel8762 3 роки тому

    Thanks a lot for this wonderful tutorial

  • @aysehamide4651
    @aysehamide4651 3 роки тому

    It's really informative. Thank you

  • @shilpasadwal2850
    @shilpasadwal2850 6 років тому +2

    Sir, can you please tell how we can take O.D.( optical density of RNA) of isolated RNA ?

    • @charleszhou5021
      @charleszhou5021 4 роки тому

      hope we can discuss further ,my email:Zgzhou@hebeismart.com

  • @michelleborrero6701
    @michelleborrero6701 3 роки тому

    Excellent! I will use it with my students.

  • @jessicanorcott7616
    @jessicanorcott7616 4 роки тому

    love the cartoon visuals !

  • @n.samantha6063
    @n.samantha6063 3 роки тому

    Thank you for your help 🙏

  • @ranirurandunu5235
    @ranirurandunu5235 4 роки тому

    Appreciate well-explained technique

  • @azif116003
    @azif116003 4 роки тому

    Yea... Very much informative. Thankyou

  • @mitylene_bailey
    @mitylene_bailey 8 років тому +1

    great! I have also seen that after the resuspension in DEPC water there is a 10-15 minute incubation at 55-60 degrees. Is that really necessary?

    • @ChangBenson
      @ChangBenson  8 років тому +1

      I believe the heating to 55-60C step you are referring to is simply to assist in resuspension of the RNA pellet because depending on the amount of RNA you have in your pellet, there may be difficulty in dissolving the RNA into solution in water.

    • @ChangBenson
      @ChangBenson  8 років тому

      I believe the heating to 55-60C step you are referring to is simply to assist in resuspension of the RNA pellet because depending on the amount of RNA you have in your pellet, there may be difficulty in dissolving the RNA into solution in water.

    • @mitylene_bailey
      @mitylene_bailey 8 років тому

      I see, thanks for the swift reply!

    • @권수현-d8f
      @권수현-d8f 8 років тому

      ChangBenson

  • @mikey27
    @mikey27 2 роки тому

    Thanks! Very useful!

  • @anisonlin6326
    @anisonlin6326 8 років тому +2

    very useful, thank you.

  • @bluespy8570
    @bluespy8570 7 років тому

    Thank you for the very useful video.

  • @buharimuhammad5989
    @buharimuhammad5989 6 років тому

    A very informative video tutorial.

  • @berry.x9388
    @berry.x9388 4 роки тому

    Thanks a lot! It was really helpful

  • @hojungyoon8239
    @hojungyoon8239 5 років тому +1

    Thank you!

  • @vijaykumar-bp4xb
    @vijaykumar-bp4xb 4 роки тому

    please mention amount of ip, trizol and other ......

  • @sakhileking6695
    @sakhileking6695 4 роки тому

    I have no idea what this is but it's very interesting.

  • @monikaantil7352
    @monikaantil7352 3 роки тому

    Very informative....thank you

  • @baitulislam3451
    @baitulislam3451 3 роки тому

    it's a great tutorial thanks a lot

  • @FrugalCreator
    @FrugalCreator 3 роки тому

    Nicely done ! Very helpful . Can you do more such lab based experimental videos ?

  • @nayimpaul3837
    @nayimpaul3837 4 роки тому

    AMAZING. THANK YOU SO MUCH.

  • @laurab1105
    @laurab1105 6 років тому

    I wish our trizol was red! Ours is yellow so it is much harder to see the difference in phase layers :(

  • @muhammadsaadansari
    @muhammadsaadansari 4 роки тому

    Very good explanation.

  • @calmrelax4447
    @calmrelax4447 4 роки тому

    awesome video. thanks!

  • @dewHong
    @dewHong 7 років тому +1

    !! it's very helpful!!

  • @leonardoguedes8817
    @leonardoguedes8817 3 роки тому

    Doesn't to be on ice?

  • @IkramUllah-ck9fi
    @IkramUllah-ck9fi 2 роки тому

    Exactly it's much more informative

  • @khubujan2369
    @khubujan2369 5 років тому

    Among the favorite ones 👍👍👍

  • @munansangu
    @munansangu 8 років тому +1

    nice video very informative

  • @paolaurbaez4312
    @paolaurbaez4312 7 років тому

    Where are the captions?

  • @Gerrahard2
    @Gerrahard2 4 роки тому

    thanks ! it was useful!

  • @ezra47986
    @ezra47986 8 років тому +1

    thank you

  • @franciscaandreamoralesherr3412
    @franciscaandreamoralesherr3412 4 роки тому

    te amo, me salvaste el pellejo muak!

  • @kunqiwang9736
    @kunqiwang9736 6 років тому

    Thanks so much.

  • @wulandariwahyuari7954
    @wulandariwahyuari7954 4 роки тому

    Thankyou for the video..

  • @mohdshahmihakimibinmazlish8914

    Can i freeze cell in -20 Celcius for 8 hours, then into liquid nitrogen for 3 months before i do the RNA extraction? anyone can help me on this?

    • @a.s9509
      @a.s9509 Рік тому

      No. You have to immediately put the samples in liquid nitrogen

  • @nancyarora7455
    @nancyarora7455 7 років тому +1

    how do we maintain pH 4 in this extraction?

    • @ChangBenson
      @ChangBenson  7 років тому

      Sorry for the delayed reply. The TRIzol reagent should already be maintained at a low acidic pH (~4), thus you should not have to worry about the pH during the extraction process. If you are encountering issues with the pH of your TRIzol reagent, you may have to either get a new bottle or make your own "TRI" guanidium-acid-phenol reagent (protocol here: www.openwetware.org/wiki/RNA_extraction_using_self-made_guanidinium-acid-phenol_reagents).
      The low acidic pH of the TRIzol reagent (~4) is essential, or else you will get both DNA and RNA in your aqueous phase.

  • @inhtung210
    @inhtung210 4 роки тому

    thanks!

  • @akashdas3003
    @akashdas3003 4 місяці тому

    Very helpful

  • @hojatabasi
    @hojatabasi 9 років тому

    thanks a lot, wery helpfull

  • @geetarani2875
    @geetarani2875 5 років тому

    so good n thanks

  • @زينزين-ط1و
    @زينزين-ط1و 4 роки тому

    Thank you so match

  • @madfolks6604
    @madfolks6604 Рік тому

    incredible

  • @ronglingwang5448
    @ronglingwang5448 9 років тому

    very good!

  • @Toretto-de-toretto
    @Toretto-de-toretto 4 роки тому +1

    masonic pyramid

  • @gaurichaudhari1848
    @gaurichaudhari1848 3 роки тому

    how to make equipment RNase free

  • @rosaceliapoquita-du8262
    @rosaceliapoquita-du8262 9 років тому +1

    thank you for this video

  • @allouchelynda5431
    @allouchelynda5431 9 років тому

    very informative. Thanks

  • @elizabethcamilapairasanchu4621
    @elizabethcamilapairasanchu4621 2 роки тому

    un traductor al español porfa

  • @ritumishra362
    @ritumishra362 8 років тому

    Bk to दर हूं धनं धन

  • @wakeup6847
    @wakeup6847 7 років тому

    it 12g, also showed 12 rcf on the centrifuge. NOT 12000G!

    • @davidmusoke
      @davidmusoke 7 років тому

      Wrong! It's 12,000g in his case. The centrifuges in our labs spin at 11,000g.

    • @wakeup6847
      @wakeup6847 7 років тому

      David Musoke wow that's so amazing. Ur centrifuge can create a black hole.

    • @davidmusoke
      @davidmusoke 7 років тому

      See the first results item after you google "RNA extraction centrifuge speed". Or look at item 11 from
      www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=2&cad=rja&uact=8&ved=0ahUKEwjiuI2Q9__XAhXIkOAKHb6DB1UQFggrMAE&url=https%3A%2F%2Fwww.scripps.edu%2Fcalifornia%2Fresearch%2Fdna-array%2Fpdfs%2FRNA%2520Extraction%2520Protocol.pdf&usg=AOvVaw3YtdKDVTLN4PRPlhDTXmAV

    • @wakeup6847
      @wakeup6847 7 років тому

      g=(1.18*10^-5)RPM.

    • @davidmusoke
      @davidmusoke 7 років тому

      A more accurate equation is g = (1.118 × 10^-5)*R* S^2, where R = radius of the spinning rotor and S = centrifuge speed measured in revolutions per minute (RPM). g is the local gravity this spinning produces. SO when a centrifuge is spec'd at 12,000g, it's an accurate representation of the amount of acceleration the RNA solution is experiencing.

  • @shenghuozhiyisi4577
    @shenghuozhiyisi4577 5 років тому

    wow!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!111

  • @tonylakouetene7239
    @tonylakouetene7239 3 роки тому

    A very informative video tutorial.