Gabatarwa:

RNA, sunan kimiyya ribonucleic acid, gajarta ta zo daga RiboNucleic Acid, wani nau'i ne na nucleic acid da aka kafa ta hanyar haɗa akalla dozin na ribonucleotides ta hanyar haɗin phosphodiester.

Tushen binciken RNA shine a fitar da jimlar RNA mara gurɓatacce, kuma ainihin ƙa'idar hakar RNA ana iya taƙaitawa kamar haka:

1. Rushe sel da farko

2. Sa'an nan kuma cire macromolecular ƙazanta (protein, lipids, DNA, da dai sauransu).

3. A ƙarshe, RNA yana haɗewa yayin cire gishiri da abubuwan da ake amfani da su, kuma yana ƙara tsarkake RNA.

Da da na yanzu na hanyar hakar

Farkon ƙarni na hanyar hakar RNA shine hanyar hazo na TRIzol (reagents da aka yi amfani da su sun haɗa da guanidine isothiocyanate, phenol, chloroform, da dai sauransu) wanda wani kakan mai suna Piotr Chomczynski da kaka Nicoletta Sacchi suka ƙirƙira, saboda wannan ƙirƙira tana ba da damar RNA cirewa cikin sauri da sauƙi, tun daga lokacin da aka fara haɓakar RNA.

Daga baya, Grandpa ya sayar da wannan haƙƙin mallaka ga Invitrogen, kuma Invitrogen ya haɓaka wannan hanya ga duniya kuma ya zama hanyar da ake amfani da ita sosai.

Daga baya, Thermo ya samu Invitrogen!

Samfurin wakilci: Invitrogen's TRIzol reagent

Koyaya, hanyar hazo na TRIzol yana buƙatar hazo da yawa da hakar RNA, kuma reagents da ake amfani da su suna da guba kuma suna ɗaukar lokaci mai tsawo don aiki.

Sabili da haka, hanyar tsarkakewar shafi-enzyme na ƙarni na biyu ya samo asali, ta amfani da silica gel membrane a matsayin takamaiman kayan tallan nucleic acid don haɓaka dawo da RNA a cikin samfurin yayin cire sauran ƙazanta.

Hanya na ƙarni na biyu yana sauƙaƙa aiki akan ingantaccen aiki, yana rage haɗarin haɗari, kuma yana haɓaka haɓakar haɓakar RNA.

Samfuran wakilai: QIAGEN RNeasy da yawancin samfuran gida

Duk da haka, duka hanyoyin ƙarni na farko da na biyu suna buƙatar amfani da DNA don cire gurɓataccen DNA, wanda ke haifar da rashin jin daɗi ga gwaje-gwajen ƙasa na gaba.Sauran DNAse zai rage ingantaccen juzu'i da kuma rage haɗin cDNA na farko, ta haka yana shafar daidaiton sakamakon.

Don haka, ana inganta hanyar haɓaka ƙarni na uku akan tushen ƙarni na biyu.Gwajin baya buƙatar ƙara kowane enzymes, kuma ana iya samun RNA cikin sauƙi tare da ginshiƙai biyu kawai.

Rukunin 1: Rukunin Tsabtace DNA don share DNA

Rukuni na 2: Rukunin RNA-kawai musamman yana haɓakawa kuma yana dawo da RNA

Irin wannan aiki mai sauƙi ba kawai yana adana lokaci ba, amma har ma yana sauƙaƙe aikin gwaje-gwaje na ƙasa!

Samfuran wakilai: QIAGEN RNeasy Plus daForegeneRNA tsarkakewa jerin kayayyakin

(RNeasy Plus)

(Foregene'sTsirrai Jimlar RNA keɓewar Kit ɗin)

Nace muku shiru!Kit ɗin RNA na Foregene shima yana gabatar da madaidaicin RNase Eraser mai sauƙin amfani, wanda zai iya cire 400μg na RNase gaba ɗaya akan tsayayyen ƙasa a cikin minti 1, ba tare da ƙara wani abu mai guba a jikin ɗan adam ba!

Hoto1: Taswirar kwatanta tsarin aiki

Kayayyakin Foregene na iya kammala fitar da RNA a cikin wakoki biyu kawai, kar ka sa wasu suyi kishin ci gaban gwajin ku!

Hoto2:Esakamakon gwaji kwatanta jadawalin

Zamanin farko (1: TRIzol)

Ƙarni na biyu (2: RNeasy na QIAGEN)

Ƙarni na uku (3: QIAGEN's RNeasy ƙari, 4: Foregene)

Hoto3: qPCR jadawali (girma mai lankwasa jadawali)

Zamanin farko (kore yana nufin TRIzol -, ja yana nufin TRIzol +)

Tsari na biyu (Sky blue yana nufin QIAGEN RNeasy-, ja yana nufin QIAGEN RNeasy +)

Ƙarni na uku (blue yana nufin QIAGEN RNeasy da, kore yana nufin Foregene)

(Lura: qPCR jadawali "-" yana nuna cewa ba a ƙara DNA ba kuma ba a cire gurɓataccen DNA ba. Saboda haka, CT ya fi girma kuma sakamakon ba daidai ba ne; "+" yana nuna cewa an ƙara DNA kuma an cire gurɓataccen DNA, sakamakon ya kasance daidai)

Kowane haɓaka fasaha yana nufin cewa aikin gwajin yana haɓakawa a cikin mafi sauƙi kuma mafi kyawun jagorar mai amfani.

Foregene yana nufin taimakawa ƙarin masu binciken kimiyya su adana lokacin gwaji da farashi, samun sakamako mai inganci na gwaji, da ƙirƙirar nasarorin binciken kimiyya tare.