Kwanan nan, na gano wani abu mai ban mamaki!Yawancin ƙwararrun ƙwararrun gwaji da ke kewaye da shi ba su ma san wasu mahimman abubuwan ilimin gwaji na asali ba.

Alal misali, za ku iya amsa tambayoyin nan?

Shin akwai bambanci tsakanin OD260 da A260?Menene kowanne yake nufi?
OD shine raguwar ƙarancin gani (yawancin gani), A shine taƙaitawar sha (sha), ra'ayoyi guda biyu a zahiri iri ɗaya ne, "yawancin gani" shine "sha", amma "yawan gani" yana cikin layi tare da mafi yawan ka'idodin ƙasa kuma mafi daidaitacce.

Yawancin lokaci muna auna ƙimar OD a 260nm don ƙididdige ƙwayar nucleic acid, don haka menene 1OD yake wakilta?
Nucleic acid yana da matsakaicin matsakaicin tsayin daka na 260nm, wanda ya ƙunshi DNA da RNA, da gutsuttsuran acid nucleic (wannan shine mahimmin batu).
Ƙimar OD da aka auna a tsawon 260 nm an rubuta shi azaman OD260.Idan samfurin ya kasance mai tsabta, ƙimar OD260 na iya ƙididdige ƙaddamar da samfurin nucleic acid.
1 OD260 = 50 μg/ml dsDNA (DNA mai igiya biyu)
= 37 μg/ml ssDNA (DNA mai ɗauri ɗaya)
= 40 μg/ml RNA
= 30 μg/ml dNTPs (oligonucleotides)
Shin akwai wata alaƙa da bambanci tsakanin RT-PCR, Realtime-PCR da QPCR?
RT-PCR gajere ne don PCR Rubutun Juya
Real Time PCR = qPCR, gajere don PCR Real Time Quantitative
Ko da yake Real Time PCR (real-time fluorescent quantitative PCR) da Reverse Transcription PCR (PCR na juye juye) duka suna da alama an taƙaita su azaman RT-PCR.Amma yarjejeniyar kasa da kasa ita ce: RT-PCR musamman tana nufin PCR na juyar da rubutu.

Menene nt, bp, da kb da aka saba amfani dasu don bayyana tsawon DNA/RNA a ilmin halitta?
nt = nucleotide
bp = tushe biyu tushe biyu
kb = kilobase

Tabbas, za ku ce mutane da yawa ba su damu da waɗannan ƙananan bayanai ba!Kowa yana yin wannan, kuma ba wanda zai tambaye ku menene.Kun san wannan ba lallai ba ne, ko?

A'a, A'a, A'a, yana da matukar muhimmanci a san wannan!saboda wanene?
Domin kuna son buga labarin!Dan uwa!Ko kuna burin kammala karatun ne ko kuna neman nasarorin binciken kimiyya, dole ne ku dogara da labaran da kuke magana!

Ya kamata hakar acid nucleic ya zama gwaji mafi sauƙi kuma mafi mahimmanci.Ingancin hakar acid nucleic kai tsaye yana ƙayyade sakamakon gwaje-gwajen da suka biyo baya.

Ko da yake na sha faɗin hakan, har yanzu akwai abokai da yawa waɗanda ba su damu ba.Wannan lokacin na yanke shawarar fita daga labarin!

Mafi ƙanƙancin Bayani don Buga Gwajin PCR na ainihin-lokaci, wanda ake kira MIQE, wani tsari ne na jagororin gwaji na kididdigar haske da aka ƙaddamar a duniya, wanda ke ba da ƙayyadaddun ƙayyadaddun ƙa'idodi don bayanan gwaji da suka wajaba don kimanta gwaje-gwajen PCR na fluorescence mai ƙididdigewa da buga labarai.Ta hanyar yanayin gwaji da hanyoyin bincike da mai gwaji ya bayar, masu dubawa za su iya kimanta ingancin tsarin gwaji na mai binciken.

Ana iya ganin cewa a cikin sashin hakar nucleic acid, an gabatar da abubuwan gano abubuwa masu zuwa.

"E" yana nuna bayanan da dole ne a bayar kuma "D" yana nuna bayanan da ya kamata a bayar idan ya cancanta.

Tsarin yana da rikitarwa sosai, a gaskiya, ina so in faɗi cewa kowa yana buƙatar farawa daga

tsarki (D), yawan amfanin ƙasa (D), mutunci (E) da daidaito (E) don kimanta acid ɗin Nucleic a cikin waɗannan abubuwa guda huɗu.

Bisa ga halaye na gwaji, da farko magana game da hanyoyin kimantawa na tsabta da maida hankali.

Ma'aunin OD shine mafi so kuma mafi sauƙi hanyar ganowa ga masu gwaji.Dangane da ka'ida, ba zan yi cikakken bayani anan ba.Yawancin dakunan gwaje-gwaje yanzu suna amfani da ultra-micro spectrophotometers don tantance samfuran acid nucleic kai tsaye.Yayin nuna ƙimar sha, shirin yana ba da ƙimar maida hankali kai tsaye (nucleic acid, furotin da rini mai kyalli) da ma'auni masu alaƙa.Amma game da nazarin ƙimar OD, Ajiye wannan hoton kuma za ku kasance lafiya.

Lissafin ƙimar ƙimar OD na duniya

Koyaya, akwai ƴan fa'idodi waɗanda ke buƙatar fitar da su daban a gare ku.

(Bayan haka, na san dole ne ku kasance masu yin ajiyar kuɗi kuma ku jira har sai kun buƙaci su!)

Bayanan kula 1 Kayan aiki

Ƙimar OD za ta shafi kayan aiki daban-daban.Muddin OD260 yana cikin takamaiman kewayon, ƙimar OD230 da OD280 suna da ma'ana.Misali, kewayon sha na Eppendorf D30 na kowa a 260nm shine 0 ~ 3A, kuma NanoDrop Daya na Thermo yana a 260nm.Matsakaicin abin sha na 0.5 ~ 62.5A.

Bayanan kula 2Dilution reagent

Ana iya shafar ƙimar OD ta hanyar dilution na reagents daban-daban.Misali, karatun OD260/280 na tsaftataccen RNA a cikin pH7.510mm Trisbuffer yana tsakanin 1.9-2.1, yayin cikitsaka tsaki mai ruwa bayanirabon zai zama ƙasa, watakila kawai 1.8-2.0, amma wannan baya nufin cewa ingancin RNA ya canza Bambanci.

Bayanan kula 3Ragowar abubuwa

Kasancewar abubuwan da suka rage za su shafi daidaiton ma'aunin maida hankali na nucleic acid, don haka ya zama dole don guje wa furotin, phenol, polysaccharide da ragowar polyphenol a cikin samfuran nucleic acid gwargwadon yiwuwa.

Koyaya, a zahiri, hakar tare da reagents na halitta tsohuwar hanya ce.A cikin kits na kasuwanci, ana iya samun tasirin hakar ta hanyar silica na tushen adsorption shafi hade tare da centrifugation, guje wa mai guba da cutarwa kwayoyin halitta waɗanda ke da wahalar cirewa, da dai sauransu Matsalar, kamar su.Foregene's nucleic acid ton kit, baya amfani da DNase/RNase da masu guba Organic reagents a duk lokacin aiki, cikin sauri da aminci., da kumatasiri shinemai kyau(A kwatsam ya ce gashi, amma na san kana son sani).

Misali 1: Yawan hakowar DNA na Genomic da tsarki

Foregene Soil DNA Isolation Kit (DE-05511) yana kula da samfuran ƙasa daga maɓuɓɓuka daban-daban, kuma adadin da tsabtar DNA ɗin da aka samu ana nuna su a cikin tebur mai zuwa:
Misali na 2: Samuwar hakar nama RNA da tsarki

Dabbobin Total RNA keɓewar Kit (RE-03012) ya sarrafa samfuran nama iri-iri, kuma adadin da tsarkin RNA da aka samu ana nuna su a cikin teburin da ke ƙasa (don ƙwayar linzamin kwamfuta):
Koyaya, kada kuyi tunanin cewa kun gama da ƙimar OD.Kuna da wani kula da mahimman abubuwan da na zana muku a gaba?

Sanarwa

Hakanan za'a ƙididdige ƙwayoyin ƙwayoyin acid nucleic a cikin abin sha.Da ɗaukan cewa kuna da ragowar DNA na kwayoyin halitta a cikin RNA, ƙimar OD ɗin ku za ta bayyana tana da girma sosai, amma ba za a iya tantance ainihin abin da ke tattare da RNA ba.Ko RNA naku Ba a bayyana ko akwai lalacewa ba, don haka har yanzu muna buƙatar cikakkiyar hanyar kimantawa don ba da ƙarin ingantacciyar hukunci, wato, ƙimar ƙimar amincin nucleic acid da aka ambata a cikin MIQE.