I-Hielscher Ultrasound Technology

I-Ultrasonic Protein Extraction kusuka ku-Tissue and Cell Cultures

  • Ukukhipha amaprotheni kuyisinyathelo esibalulekile sokulungiselela isampula kuma-proteomics.
  • Amaprotheni angathathwa ezilwaneni zezitshalo nezesilwane, ama-yeasts nama-microorganisms.
  • I-Sonication iyindlela enokwethenjelwa, ephumelelayo yeprotheyini yendlela yokwenza amaprotheni aphezulu avele ngesikhathi esifushane sokukhipha.

 

Ukukhishwa kwamaphrotheni kusuka ezicutshini nasemangqamuzaneni

Izakhi zamaprotheni ezivela emathisini namaseli akhiqizwa kuyisinyathelo esibalulekile sokulungiselela isampula esenziwa ngesikhathi samasu amaningi wezinto eziphilayo kanye nokuhlaziya ezifana ne-ELISA, IKHASI, I-Western blotting, mass spectrometry, noma amaprotheni ukuhlanzwa. I-Ultrasonic cell ukuphazanyiswa, lysis kanye nokukhipha iyindlela ephathekayo efanelekayo ukuqinisekisa ukuvunwa okuphezulu kwamaprotheni.

Amaprotheni isisindo esivela ezilwaneni zesilwane

Ukuze kulungiswe izicubu zomzimba wonke (isib. Izinso, inhliziyo, amaphaphu, imisipha njll), izicubu kufanele zihanjiswe ezingxenyeni ezincane kakhulu ngamathuluzi ahlanzekile, ngeqhwa ngokukhethekileyo, futhi ngokushesha ngangokunokwenzeka ukuvimbela ukuchithwa ngamaprotheni (isib. i-lysis buffer efana ne-RIPA noma i-hypotonic lysis buffer equkethe i-protease ne-phosphatase inhibitor cocktail). Ngemuva kokusabalalisa, isampula igxila ekwenzeni i-nitrogen enamandla ukuze iqhwaze. Isampula ingagcinwa ku -80 ° C ukuze isetshenziswe kamuva noma ihlale iqhwa ukuze i-homogenization esheshayo. Ngokushesha ngaphambi kokukhipha i-ultrasonic, i-ice cool lysis buffer (ne-protease inhibitors DTT, leupeptin ne-aprotinin) ifakwe ngokushesha kwi-tube yesampula (ngayinye ye-tissue approx. Okungenani. 20-60mg wezincubu kunconywa nge-tube yesampula ngayinye.
I-ultrasonic homogenization, i-lysis kanye ne-extraction yenziwa nge-homogenizer ye-ultrasonic efana ne- UP200Ht noma UP200St, ifakwe ne-sonotrode encane. Isikhathi se-Sonication singama-60-90 sec. kwimodi ye-ultrasonic yomjikelezo we-15 sec. sonication kanye 10 sec. isikhathi sokuphumula. Isampula kufanele igcinwe eqhweni ngaso sonke isikhathi.
Ngemuva kwe-homogenization ye-ultrasonic / isizinda, i-lysate i-centrifuged ku-27,000g nge-approx. 20 iminithi. Emva kwalokho i-supernatent iqoqwe, ukuze ukuhlushwa kwamaprotheni kunqunywe yi-protein test ezifana nePyerce protein testay BCA.

I-sonication yamaprotheni iyindlela ebalulekile yokulungiswa kwesampula

Ukukhishwa kweprotheni ye-Ultrasonic kusuka kumaseli nge UP200St

Isicelo solwazi




Qaphela kwethu Inqubomgomo yobumfihlo.


Iprotheyini yesikhumba kusuka ku-serum yegazi

Ukuze uthole ingxube ehambisanayo ye-serum kanye ne-phosphate buffer, isampula ihamba phambili ngaphambi kwe-ultrasonic cell lysis. Ukuze i-ultrasonic lysis, isampula i-sonicated ne-ultrasonic lab homogenizer efana ne- UP100H imijikelezo engu-8 ku-20% amplitude, ngokuhamba kwemizuzwana ngayinye emihlanu nangemizuzwana engu-15. I-Sonocation iqhutshwa yi-sonicationg emjikelezweni futhi ngokubeka isampula eqhweni ukuze ukugwema ukushisa nokushisa kwesampuli kugwenywe. Njengoba i-serum iqukethe inani elikhulu lamaprotheni ezinyathelo ezinamandla (njenge-albin, i-α1-antitrypsin, i-transferrin, i-haptoglobulin, i-immunoglobulin G kanye ne-immunoglobulin A), ephazamisa ukuhlukaniswa kwamaprotheni aphansi kwamangqamuzana e-IEF, kunconywa ukuba ichithe kusuka ku-serum usebenzisa ikholomu yokunciphisa.

Amaprotheni isisindo esivela emathini esitshalo

Amasha, izitshalo ezithambile ezithambile, isib. Izinyosi njll, zingaphazanyiswa kalula ngokufaka nje isampula eqoshiwe e-lysis buffer for sonication. Ama-tough, ama-tissue ezimila ezimila, njengezimbewu, izinaliti ezinamathele njll, kufanele zibe nomhlabathi omile. Ezinye izitshalo eziqinile, ezinomsoco kumele zifakwe efrijini futhi zibe semhlabathini e-nitrogen elamanzi ngaphambi kokukhishwa nge-sonication. Ukumiswa kwamasiko esitokisi wesitshalo, ukwelashwa kwe-ultrasonic emkhatsini wamasekhondi angu-30 no-150 ku-buffer buffer ikakhulukazi ngokwanele. Izinto eziphathekayo ezinjengezimpande zifuna i-sonication ekhudlwana njengoba kuchaziwe ngezansi.

I-Protocol ye-ultrasonic isizinda se-albinini kusuka kumbewu yamathanga

I-ultrasonic izicubu ze-homogenizer UP400St ene-S24d40 - yeprotheyini yesikhumba kusuka kwezitshalo nezilwane zesilwane (Chofoza ukuze ukhulise!)Ukuze uthole i-ultrasonic protein extraction ye-albinini kusuka emhlabathini omuhle we-seed powder, u-10 g we-pumpkin we-seed powder ohlukanisiwe kanye namamitha ayi-100 wamanzi abiziwe njengamakhemikhali afakwe engxenyeni engu-250mL ye-glass beaker. Isiqephu seprotheyini sinezinyathelo ezimbili: Okokuqala, isampula ihanjiswa nge-probe-type ultrasonicator I-UP400St (400W, 24kHz) nge sonotrode ehlanganisiwe S24d7. I-beaker yeglasi ifakwa emanzini okugeza abandayo ngesikhathi se-homogenization ye-ultrasonic. Ukubekwa kwe-ultrasonicator UP400St ne-sensor yokushisa ekhuphukayo yaqinisekisa ukuthi izinga lokushisa lwesampula lihlale ligcinwe ngaphansi kwe-30 ° C. Ngokulawulwa kokushisa okuqondile ngesikhathi sonication, i-denaturation ye-albinin igwenywe. Okwesibili, ukukhishwa kwenziwa nge-mixer ngo-200 rpm ngesivinini futhi ku-30 ​​° C. Ngemuva kwalokho i-beaker idluliselwa ku-shaker therkeratic. I-globulin isuswa nge-dialysis ngamanzi a-distilled. Ngemuva kokukhishwa kwe-globulin, ukukhishwa kwamaprotheni kungathathwa isampuli ngokuzimisela kwephrofayela ye-albin bese kulungiswa ku-pI = 3.0 besebenzisa iHCl eyi-0.1 M ye-albin coagulation. Isigaba esiqinile sihlukaniswa yi-centrifugation ku-5000g, 20 ° C futhi ibuye ibuye ibuyele emanzini amisiwe. I-albin coagulation yenziwa kabili ukwandisa isilinganiso seprotheyini ku-concentrating albinini.

Ultrasonic alkaline amaprotheni isizinda sokulungiselela amaprotheni ukugxila kusuka ilayisi bran kubonisa ukuthi ukwelashwa ultrasonic kuholela amaprotheni ephakeme isivuno esikhathini esifushane kakhulu isizinda isikhathi – uma kuqhathaniswa nezindlela ezivamile zokufakelwa.

Idivayisi ye-ultrasonic i-VialTweeter ye-protein isakhiwo kusuka kumasampula wezicubu (Chofoza ukuze ukhulise!)

I-VialTweeter for sonication ngqo.

Izinzuzo

  • ngokushesha
  • izithelo eziphakeme
  • ephumelelayo kakhulu
  • Ukulawula okucacile mayelana nemingcele
  • imiphumela ye-reproducible
  • ukusabalalisa okulinganayo

Isampula yokulungiselela isimiso se-enzyme ye-iNOS esebenzayo

Ukuze uthole i-enzyme ye-iNOS esebenzayo ngokugcwele (isib. Ukuhlolwa kwezidakamizwa), le protocol elandelayo inconywa: ukumiswa kweseli kufanele kufakwe kwiqhwa bese kukhulunywa nge-an UP100H ku-10μm amplitude kumodi yokujikeleza ka-5 sec. sonication kanye 25 sec. uhlale phezu kweqhwa. Inqubo kufanele iphindwe ngokuphindaphindiwe. izikhathi ezingu-3. Isikhathi sokuphumula phakathi kwemijikelezo ye-sonication sinciphisa izinga lokushisa futhi ngaleyo ndlela lizonciphisa ingozi yokuqothula.

I-Ultrasonic Protein Solubilization

I-Sonication ingase ijubane nenqubo ye-protein yokuxazulula, okuvame ukudinga amahora amaningana. Ukuze ungadluli isampula futhi uvimbele ukuchithwa kwamaprotheni nokuguqulwa kwezixazululo eziqukethe i-urea, ukuqhuma kwe-ultrasonic akufanele kudlule isikhathi eside kunamasekhondi ambalwa.

Imiyalo Ejwayelekile

Ukulawula ukushisa: Ukuze kuqinisekiswe ukuthi amaprotheni aphezulu akhiqizwa ngaphandle kwe-denaturation eshisayo, izinga lokushisa ngesikhathi sokukhipha kumele kulawulwe. I-homogenizers ye-Hielscher ye-state-of-art ultrasonic – futhi ubizwa ngokuthi i-ultrasonic disintegrator noma i-sonificator – zilawulwa ngokuqondile. Bafika ngenenjini yokushisa eguquguqukayo. Ezokukhethwa kukho kokuhlelwa kwe-homogenizer ye-ultrasonic, izinga lokushisa eliphezulu lingasethwa. Uma le max yokushisa isifikile, i-ultrasonicator ivele ivele ize isetshenziswe phansi.
I-Buffer: Ukukhethwa kwesiphakamiso esifanelekile kanye nevolumu efanele ye-buffer kuyahlukahluka kusuka kumathishu kuya kwezicubu futhi kufanele kucatshangwe-ukuhlolwa kokuhlolwa kwesilingo nokuphutha.
Ukuhlukaniswa / ukuhlanzwa: Ama-protein lysates angaqukatha ama-biomolecules amaningi ngokweqile njenge-DNA noma ama-carbohydrate, angase asuswe yi-protein precipitation (deoxycholate-trichloroacetic acid) noma ukushintshaniswa kwe-buffer.

I-Chittapalo noNoomhorm (2009) yabika ukuthi amaprotheni akhiqiza ukwanda ngokusebenzisa i-sonication nokuthi inqubo ye-ultrasonic ye-tissue homogenization kanye nenqubo ye-lysis ingakwazi ukwandisa izinqubo zokukhipha ezikhona kakhulu – ukuvumela amathuba amasha wokungenisa ezentengiselwano.

Ukuvikelwa komsindo nge-enclosure ye-Hielscher ye-SPB-L nedivayisi ye-ultrasonic UP200St. (Chofoza ukuze ukhulise!)

Ultrasonic izicubu ze-homogenizer UP200St ukuze usebenzise kahle amaprotheni

Ukulawula okucacile ukwelashwa kwe-ultrasonic (Chofoza ukuze ukhulise!)

Ukulawulwa kwesiphequluli sokusebenza okuqondile nokuqapha inqubo ye-sonication

Izinsiza ze-Ultrasonic zeProtein Extraction

I-Hielscher Ultrasonics inikeza ububanzi obubanzi be-homogenizers e-ultrasonic ngokuhlukaniswa kwamangqamuzana, izicubu, amabhaktheriya, ama-microorganisms, imvubelo nezinhlamvu.
I-Hielscher lab ultrasonicators inamandla futhi kulula ukuyisebenzisa. Eyakhelwe ukusebenza kwe-24/7, yenzelwe njenge-lab enamandla futhi ephumelelayo namadivaysi phezulu. Kuzo zonke amadivaysi, ukukhishwa kwamandla kanye ne-amplitude kungalawulwa ngokuqondile. Ububanzi obanzi Izesekeli ivula izinketho ezengeziwe zokusetha. Amadivayisi digital ezifana I-VialTweeter, UP200Ht, UP200St, futhi UP400St babe nokulawula okushisa okudidiyelwe kanye nekhadi le-SD elakhiwe ngokuzenzakalelayo lokurekhoda kwedatha.
Ngokungaqondile, ukungahambisani nokungcola-kanye nokunikezwa kwe-sampuli amaningi ngesikhathi esisodwa, sinikela I-VialTweeter noma i I-ultrasonic cuphorn.
Kuye ngesicelo sakho, izinto ezibonakalayo, nesampuli yesampula, sizokuncoma ukuthi usethe uhlelo oluhle kakhulu lwesampula yakho yesampula. Xhumana nathi namuhla!

Xhumana nathi! / Cela Us!

Sicela usebenzise ifomu elingezansi, uma ufisa ukucela ulwazi oluthe xaxa mayelana ne-homogenization yama-ultrasonic. Sizojabula ukukunikeza uhlelo lwe-ultrasonic ukuhlangabezana nezidingo zakho.









Sicela uqaphele wethu Inqubomgomo yobumfihlo.


Izincwadi / Izinkomba

  • I-Chittapalo T, i-Noomhorm A (2009): I-Ultrasonic yasiza ukukhipha ama-alkali kwiprotheni kusuka ku-rice bran nesakhiwo seprotheyini sigxila. Int J Food Sci Technol 44: 1843-1849.
  • Simões, André ES:; Pereira, uDiane M .; I-Amaral, uJoana D.; I-Nunes, i-Ana F .; Gomes, Sofia E .; Rodrigues, uPedro M .; Bheka, Adrian C .; I-D'Hooge, i-Rudi; I-Steer, uClifford J .; UThibodeau, uStephen N .; Borralho, Pedro M .; U-Rodrigues, i-Cecília MP (2013): Ukuphumula okuphumelelayo kwamaprotheni avela kumasampula amaningi emithombo emva kwe-trizol noma i-trizol LS i-RNA kanye nesitoreji sesikhathi eside. BMC Genomics 2013, 14: 181.


Amaqiniso Okufanele Ukwazi

Ama-proteomics

I-proteomics yinkambu yokucwaninga ehlola amaprotheni kanye neproteome. Ama-proteins agcwele uhlaka oluningi lwemisebenzi ebalulekile ngaphakathi kwezinto eziphilayo. I-proteome yilo lonke iqoqo lamaprotheni elivezwe yi-genome, iseli, izicubu, noma imvelo ngesikhathi esithile. I-proteome ihluka ngesikhathi kanye nezidingo ezihlukile, noma ukucindezeleka, ukuthi iseli noma imvelo ihamba. Ngokuqondile, kuyisethi yamaprotheni abonisiwe ngohlobo olunikeziwe lweseli noma inyama, ngesikhathi esinikeziwe, ngaphansi kwezimo ezichazwe. Leli gama liyingxube yamaprotheni ne-genome. Ama-proteomics ukutadisha i-proteome.

Amaphrotheni

Ama-protein ayimvelo ye-biomolecules, okuthiwa i-macromolecules – okuyinto eyenziwa kusuka eminye imikhakha emide yama-amino acid. Ama-proteins akhona kuzo zonke izilwane zombili zemifino nezilwane futhi zibaluleke kakhulu emisebenzini eminingi yezinto eziphilayo. Njengoba amaprotheni aqukethe ulwazi oluningi lwezinto eziphilayo, athathwa ngenhloso yokuhlaziya, isib. Ukucwaninga kwe-proteomic. Umsebenzi obaluleke kunazo zonke owenziwe amaprotheni kufaka ukucubungula kokusabela kwamagciwane, ukuphindaphinda kwe-DNA, ukuphendula okungahambi kahle, nokuthuthwa kwama-molecule kusuka endaweni eyodwa kuya kwenye. Amaprotheni ahluke kakhulu ngokulandelana kwawo kwama-amino acids, okushiwo ukulandelana kwe-nucleotide yezinhlobo zawo, futhi okuvame ukuphumela kumaprotheni ukuphoqa kwisakhiwo esithile esilandelayo esinqumayo umsebenzi wawo. Ama-proteine ​​ayenjalo – ngaphandle kwama-peptide – enye yezingxenye eziyinhloko zokudla. Ngakho-ke, i-proteomics iyithuluzi elinamandla kwisayensi yokudla yokwandisa izinqubo, ukuphepha kokudla nokuhlola okunomsoco.

I-Gel Electrophoresis

I-gel electrophoresis iyindlela enkulu yokuhlukanisa nokuhlaziywa kwama-macromolecules afana ne-DNA, i-RNA namaphrotheni kanye neziqephu zazo, ngokususelwa ngobukhulu bawo kanye nokukhokha. Isetshenziswe emakhemikhali omtholampilo ukuze ahlukanise amaprotheni ngokukhokhiswa kanye / noma usayizi (IEF i-agarose, ngokujwayelekile usayizi ozimele) naku-biochemistry, i-biology yamangqamuzana kanye ne-proteomics ukuhlukanisa inani elixubile le-DNA ne-RNA fragments ubude, ukulinganisa ubukhulu be-DNA kanye ne-RNA fragments noma ukuhlukanisa amaprotheni ngokukhokhiswa.

Amasiko weselula

Isiko le-cell yiyona inqubo ekhulayo elawulwayo lapho amaseli ahlwanyelwa khona ngaphansi kwezimo ezilawulwayo. Izimo zamasiko zeselula ziyahlukahluka ngohlobo ngalunye lweseli. Ngokuvamile, imvelo yesitokisi leseli iqukethe isitsha esifanele (isb. Petri dish) nge-substrate noma ephakathi enikeza izakhi ezibalulekile (ama-amino acids, ama-carbohydrate, amavithamini, amaminerali), izimo zokukhula, ama-hormone, nama-gas (CO2, O2), futhi ilawula isimo se-physio-chemical (pH buffer, pressure osmotic, izinga lokushisa). Amangqamuzana amaningi adinga i-substrate ebusweni noma ngaphezulu, kanti amanye amasiko esitokisini angalinywa mahhala enkathini yamasiko (isiko sokumiswa, ukumiswa kweseli).
Amasiko amasiko ezinhlayiya zezilwane asetshenziselwa ukukhiqizwa kwezimboni zegciwane legciwane lesandulela ngculaza kanye neminye imikhiqizo ethathwe yi-biotechnology. Amaseli e-stem abantu ahlonyelwe ukukhulisa inani lamaseli futhi ahlukanise amaseli abe yizinhlobo ezahlukene ze-somatic cell for izinjongo zokufakelwa kabusha.

Amasampula weTishu

Lezi zicubu zichaza isisemkhatsini seselula, lapho impahla yeli cell isesimweni senhlangano phakathi kwamaseli nesitho esiphelele. Ezingxenyeni, amangqamuzana afanayo, avela emlandweni ofanayo owenza ndawonye umsebenzi othile, ahlangene. Ngokuhlanganiswa okusebenza kwamathambo amaningi, izakhiwo eziyinkimbinkimbi zezitho zakhiwa.
Izicubu zisampuliwe ekucwaningweni kwe-biology, i-histology / hertopathology, i-parasitology, i-biochemistry, i-immunohistochemistry kanye nokuhlakulela nokukhipha iDNA. Ingahlukaniswa phakathi kwesilwane (ukuhlukaniswa iziqephu: izicubu ezincelisayo) kanye nezicubu zezitshalo. Izicubu zezilwane zihlukaniswe zibe yizinhlobo ezine eziyisisekelo zezicubu ezixhumeneyo, ezinesisindo, ezesabekayo, nezase-epithelial. Izicubu zezitshalo zihlukaniswe kulezi zindlela ezintathu ezilandelayo: i-epidermis, izicubu zomhlabathi, nezicubu zesisulu.
Ama-sampuli we-tissue angalungiswa kusuka ezingxenyeni zezilwane noma zezitshalo, isib. Amathambo, imisipha, amaqabunga, njll.

Izifo zomzimba

Igazi, i-serum, i-plasma, i-cerebrospinal fluid, i-saliva ne-synovial fluid yiziqu zomzimba, ezinikeza umthombo omkhulu wokuthola ulwazi olufanelekile. Ngakho-ke, ukulungiswa okunamandla kwamasampula omzimba womzimba ukuhlaziywa kubalulekile. Inselele yokuqala ihlotshaniswa nobubanzi obanzi obukhulu bezingxenye ezitholakala emzimbeni womzimba.

Ukunqunywa kwamaprotheni ukuhlushwa

U-Bradford assay, isilingo se-Lowry nesilingo se-bicinchoninic acid (BCA) yizivivinyo ezivamile zokunquma ukuxilongwa kwamaprotheni. I-bovine serum albin (i-BSA) ingenye yezinga eliphezulu lamaprotheni asetshenziswe kakhulu.

Lysis Buffer

I-Lysis buffer kufanele ikhethwe ngokuhambisana nezinto eziselukwini noma izicubu (izicubu zesitshalo, isitshalo, amabhaktheriya, isikhunta, njll), nokuthi ngabe amangqamuzana asesakhiwo kanye nohlobo lwesakhiwo. Ama-buffers amaningi we-lysis wokukhishwa kwamaphrotheni, amakhanda, nama-organelles akhiwe nge-detergents eyodwa noma ngaphezulu. I-detergent ivame ukukhethwa ngokuhlolwa kwezivivinyo-nephutha noma – uma kutholakala – ngokusho kweprotein yamaphrotheni ekhona. I-detergent kumele ihambisane nomthombo wamathambo namaprotheni. Ngokuvamile, isilawuli esincane kakhulu esisebenzela isisindo / amaprotheni ethile, sikhethwe ukuze sigcine ukusebenza okuphezulu kokukhipha. Ngaphezu kwalokho, uma kwenzeka ukukhishwa kwamagundane kanye nama-organelles, i-detergent emnene igcina i-membrane ifinyelele. Izitshalo ezivame ukusetshenziselwa ukuphengula i-lysis ziyi-nonionic noma zwitterionic, isib. CHAPS, deoxycholate, iTriton ™ X-100, NP40, ne-Tween 20.
Ngokwesibonelo, izicubu ezifana nobuchopho, isibindi, amathumbu, izinso, isiteji njll zingamane zigcwale i-RIPA – kodwa i-protease inhibitors ne-DTT (isib. i-gel electrophoresis) kufanele ifakwe.
I-Lysis buffer yama-muscle muscle (ice frost): i-TM i-20mM (pH 7.8), i-NaCl eyi-137mM, i-KCl engu-2.7mM, i-1 mM i-MgCl2, i-1% i-Triton X-100, i- 10% (w / v) i-glycerol, i-1 mM EDTA , I-1 mM i-dithiothreitol eyongezwa nge-protease ne-phosphatase inhibitor cocktail
Ithebula lezinto ezijwayelekile zokuthenga kanye nohlobo lwe-pH. Ngokuvamile, lezi ziphazamisi zivame ukusetshenziselwa ukugxila kwezingu 20-50mM.

I-Buffer uhla lwe-pH
Citric acid – I-NaOH 2.2 – 6.5
I-citrate ye-sodium – Citric acid 3.0 – 6.2
I-acetate ye-sodium – i-acetic acid 3.6 – 5.6
Usawoti kaCacodylic acid sodium – I-HCl 5.0 – 7.4
MES – I-NaOH 5.6 – 6.8
I-sodium dihydrogen phosphate – i-disodium hydrogen phosphate 5.8 – 8.0
Imidazole – I-HCl 6.2 – 7.8
I-MOPS – KOH 6.6 – 7.8
I-triethanolamine hydrochloride – I-NaOH 6.8 – 8.8
I-Tris – I-HCl 7.0 – 9.0
HEPES – I-NaOH 7.2 – 8.2
I-Tricine – I-NaOH 7.6 – 8.6
I-tetraborate ye-sodium – i-boric acid 7.6 – 9.2
Ibhayisikili – I-NaOH 7.7 – 8.9
Glycine – I-NaOH 8.6 – 10.6

Amanothi amaningi abonisa ukuthembeka kwe-pH ngokushisa. Lokhu kuyiqiniso ikakhulukazi ku-Tris buffers. Izinguquko ze-pKa zisuka ku-8.06 ku-25 ° C kuya ku-8.85 ku-0 ° C.
(i-pH ne-pKa ye-buffer: i-pH iyalinganisa ukuhlushwa kwama-hydrogen ion kwisixazululo se-aqueous. i-pKa (= i-acid dissociation constant) isilinganiso esilandelayo, kodwa esicacile kakhulu, ngoba kusiza ukubikezela ukuthi i-molecule izokwenza kanjani ngokuqondile pH value.)

I-TRIzol

I-TRIzol iyisisombululo sekhamikhali esisetshenziselwa ukukhipha i-RNA / DNA / amaprotheni ngesikhathi se-guanidinium thiocyanate-phenol-chloroform isizinda. Ukusetshenziswa kwe-ultrasonically kusiza isizinda se-TRIzol imiphumela ku-DNA ephezulu, i-RNA, namaprotheni okukhiqizwa kwisampula esifanayo futhi ihamba phambili ngaleyo ndlela nezinye izindlela zokutshala.