nuclic acidche praman

aanvik jivashastrat, mishranat asalelya dii. aine. aii. kinwa aar. aine. aii. chee sarasari sandrata tasech tyanchi shuddhata nirdharit karnyasathi samanyatah nucleic acidche praman nishchit kele jaate. nuclic acidcha vapar vapar pratikriyansathi bahudha ishtatam kamgirisathi vishisht pramanat aanee shuddhatechi aavashyakta asate. aajparyant, dravanatil nucleic acidche (jase qee dii. aine. aii. kinwa aar. aine. aii.) praman nirdharit karnyasathi kinwa sandrata sthapit karnyasathi shastradnyankadun done mukhya paddhati vaparlya jaatat. heey dii. aine. aii. rangachya upasthitit spectrophotometric parimanankan aanee yuu. vhee. pratidipti tagging aahet.
spectrophotometric vishleshan.
dii. aine. aii. kinwa aar. aine. aii. chey praman nirdharit karnyasathi sarvat samanyapane vaparlya jaanarya paddhatinpaiki eka mhanje spectrofotometercha vapar karoon spectrophotometric vishleshanacha vapar. spectrophotometer mishranat asalelya dna kinwa rna yaa nuclic acidchi sarasari sandrata tasech tyanchi shuddhata nirdharit karnyas saksham aahe.
spectrophotometric vishleshan heey nucleic acid eka vishisht namunyat atinil prakash shoshun ghetat yaa tattvanvar aadharit aahe. dii. aine. aii. aanee aar. aine. aii. chya babatit, eka namuna 260 nanometer (aine. emm.) chya tarangalambivar atinil prakashachya samparkat yeto aanee photo-detector namunyatoon jaanara prakash mojato. kahee atinil prakash tyatoon jaeel aanee kahee dii. aine. aii./aar. aine. aii. dware shoshale jaatil. namunyane jitka jaast prakash shoshala jaeel, namunyatil nucleic acidche praman titkech jaast asel. parinami honara parinam assa aahe qee prakash shodhkavar qamee prakash padel aanee yamule uchch prakashik ghanata (oo. dii.) nirman hoil.
bear-lambert kayadyacha vapar karoon shoshalelya prakashachya pramanacha sambandh shoshak renuchya ekagrateshi jodane shakya aahe. 260 nmchya tarangalambivar, duheri-stranded dnsathi sarasari vilupta gunank 0.020 (microgram/miilii) −1 semee−1 aahe, ekal-stranded dnsathi too 0.027 (microgram/miilii) −1 semee−1 aahe, ekal-stranded rnsathi too 0.025 (microgram/miilii) −1 semee−1 aahe aanee lahan ekal-stranded oligonucleotides too lambi aanee bases rachnevar avalambun aahe. asha prakare, 1 chaa shoshak (aaa) haah duheri-adakalelya dii. aine. aii. sathee 50 microgram/milichya sandrateshi sambandhit aahe. ganana karanyachi hii paddhat kiman 2 chya aii paryant vaidh aahe. oligonucleotides adhik achook vilupta gunank aavashyak asoo shakato; jawalche shejari model vaparoon yacha andaj lavala jaau shakato.
ganana.
optical ghanata samikaranatun nirman hoteh
vyavharik drushtya, jyaa namunyat dii. aine. aii. kinwa aar. aine. aii. nasale, tyane konatyahi atinil prakashala shoshun gheu naye aanee tyamule 0 chee oo. dii. nirman karavi.
prakashik ghanata = logg (100/100) = 0
dii. aine. aii. kinwa aar. aine. aii. chey praman nirdharit karnyasathi spectrophotometric vishleshan vapartana, manak vakranchi aavashyakta naan thevata adnyat praman nirdharit karnyasathi bear-lambert kayada vaparla jaato. thodakyat, bear lambert kayada shoshun ghetalelya prakashachya pramanacha sambandh shoshak renuchya ekagrateshi jodane shakya karato. khaleel shoshak ghatak nucleic acid ekagrata rupantaran ghatakanmadhye vaparale jaatat, jeey adnyat nucleic acid namunyanchya oo. dii. chey ekagratemadhye rupantar karatatah
rupantaran ghatak.
10 miimii lambicha marg vapartana, ekagrata nirdharit karnyasathi oo. dii. laa rupantaran ghatakane gunakar qaraa. udaharanarth, 2 oo. dii. dii. ess. dii. aine. aii. chaa namuna 100 microgram/miilii sandrata asalelya namunyashi sambandhit aahe.
10 mimipeksha qamee lambicha marg vapartana, parinami oo. dii. 10/lambichya ghatakanam qamee hoil. 3 miimii lambichya margachya vareel udaharanacha vapar karoon, 100 microgram/miilii namunyasathi oo. dii. 0.6 paryant qamee kele jaeel. 10 miimii samatulya ekagrata samanya karnyasathi, khaleel goshti kelya jaatatah
0.6 od x (10/3) * 50 microgram/miilii = 100 microgram/miilii
bahutek spectrophotometer nuclic acidcha prakar aanee margachi lambi asha prakare nivadu detat qee parinami ekagrata 10 miimii margachya lambiparyant samanya hote jee beearchya kayadyachya tattvanvar aadharit aahe.
praman mojmap mhanoon a260.
nucleic esidsathi praman mojmap mhanoon "aii260 unit" vaparale jaate. eka aii260 ekak mhanje 1 milimadhye asalele aanee 1 oo. dii. tayar karnare nuclic acidche praman. samaan rupantaran ghatak lagoo hotat aanee mhanoon, asha sandarbhatah
namunyachi shuddhata (260:280/260:230 gunottar).
nucleic acidche namune itar renunmule dooshit hone samanya aahe (i.eae. prathine, sendriya sanyuge, itar). nuclic aesidchya pramananasathi spectrophotometric vishleshan vaparnyacha duyyam fayda mhanje 260 nmh 280 nm ganana vaparoon namunyachi shuddhata nirdharit karanyachi kshamata. nucleic aesidchya shuddhateche moolyankan karnyasathi 260 aanee 280 nm (aii 260/280) yaa shoshanache gunottar vaparale jaate. shuddha dii. aine. aii. sathee, 260/280 heey mothya pramanavar ~ 1.8 manale jaate parantu mool warberg peparmadhil sankhyatmak chukanmule-60 takke prathine aanee 40 takke dii. aine. aii. chya mishranat anuvadit karnyacha yuktivad keela gela aahe. shuddha aar. aine. aii. aii. 260/280 chey gunottar ~ 2 aahe. heey gunottar samanyatah nuclic acid isolation prakriyetoon uralelya prathinanchya pradushanachya pramanache moolyankan karnyasathi vaparale jaatat kaaran prathine 280 nmvar shoshun ghetat.
prathine (visheshtah, sugandhit amino aaml) 280 nmvar prakash shoshun ghetalyamule, samanyatah prathinanchya dravananchya dna pradushanache moolyankan karnyasathi 260 nm viruddha 280 nmvaril shoshanache praman vaparale jaate. matra, yachya ulat kharey nahee-nucleic acid dravanatil 260:280 gunottaravar lakshaniy parinam karnyasathi prathinanche pradushan tulanene mothya pramanat lagate.
260:280 gunottaramadhye prathinanmadhil nuclic aesidchya dushitatebaddal uchch sanvedansheelata asateh
nucleic acidemadhil prathine dooshit honyabaddal 260:280 gunottarat sanvedansheelata nasate (rnsathi darshavilele takta, 100% dna andaje 1.8 aahe):
haah farak prathinanchya tulanet nucleic aesidmadhye 260 nm aanee 280 nm itake jaast vastuman ksheen gunank asalyamule aahe. yamule, prathinanchya tulanene uchch sandratesathi dekhil, prathin 260 aanee 280 shoshanamadhye tulanene qamee yogdan dete. prathinanchya dushitateche moolyankan 260:280 gunottarane vishvasarhpane kele jaau shakat nasale taree yacha arth assa dekhil aahe qee teey dna praman andajat thodi truti yogdan dete.
dushitatechi olakh.
namuna shuddhateshi sambandhit samasya astitvat aahe heey olakhanyasathi namuna varnapat tapasane upayukt tharoo shakate.
fluorescent dye taggings vishleshan.
dii. aine. aii. aanee aar. aine. aii. chya sandrateche moolyankan karnyasathi eka paryayi paddhat mhanje namunyala fluorescent tags tagg karane, joou eka fluorescent rung aahe joou nuclic acidshee bandhalelya ranganchi tivrata mojanyasathi vaparla jaato aanee bandhalyavar nivadakpane fluoros hoto (udaa.jee. ethidium bromide). hii paddhat asha prakarananmadhye upayukt aahe jithe spectrophotometridway achook moolyankan karnyasathi ekagrata khoop qamee aahe aanee jyaa prakarananmadhye 260 nmvar dooshit padarth shoshun ghetalyane tyaa paddhatidvare achook pariman ashakya hote. dii. aine. aii. aanee aar. aine. aii. chya pratidipti parimananacha fayda mhanje spectrophotometric vishleshanapeksha sudharit sanvedansheelata. tathapi, sanvedanshilatemadhye jhaleli tee vaadh prati namuna jaast kimmat aanee namuna tayar karnyachya dirgh prakriyechya kimmativar yete.
yaa dishene jaanyasathi done mukhya marg aahet. "spotting' madhye namuna thet agroj jail kinwa plasticchya gundalanyavar thevane samavisht aahe. fluorescent rung ektar agroj jailmadhye asato kinwa plasticchya patalavaril namunyanmadhye yogya pramanat jodala jaato. namunyachya bajula dnyat sandrata asalelya namunyancha sanch aadhalato. yaa dnyat sandratechya pratidiptishi tulana karoon adnyat namunyachya sandratecha andaj lavala jaato. vaikalpikaritya, dnyat sandratechya kahee namunyansah, ekhada jaroj kinwa polyacrylamide jeldware namuna chalvu shakato. spott chachanipramanech, dnyat namunyanshi fluorescent tivratechi tulana karoon ekagratecha andaj lavala jaato.
jarr namunyanche praman microplates kinwa couvets vaparnyasathi purese mothe asel tarr rangane bharalele namune pratidiptimapkadvare dekhil mojale jaau shakatat. kiman namuna aakarman 0.3 microlpasoon suru hote.
260 aine. emm./280 aine. emm. spectrophotometric aavruttisarkhich asalelya dii. aine. aii. namunyache prathine dooshit karane nirdharit karnyasathi aajtagayat konatihi pratidipti paddhat nahee.

