Video: Yuav ua li cas DNA concentration xam nrog spectrophotometer?
2024 Tus sau: Miles Stephen | [email protected]. Kawg hloov kho: 2023-12-15 23:36
DNA concentration yog kwv yees los ntawm ntsuas qhov nqus ntawm 260nm, kho qhov A260 ntsuas rau turbidity ( ntsuas los ntawm absorbance ntawm 320nm), sib npaug los ntawm lub dilution factor, thiab siv kev sib raug zoo uas A260 ntawm 1.0 = 50µg/ml ntshiab dsDNA.
Tib neeg kuj nug, koj pom qhov concentration thiab purity ntawm DNA li cas?
Txhawm rau ntsuas DNA purity , ntsuas absorbance los ntawm 230nm mus rau 320nm txhawm rau txheeb xyuas lwm yam kab mob. Feem ntau purity xam yog qhov piv ntawm qhov nqus ntawm 260nm muab faib los ntawm kev nyeem ntawv ntawm 280nm. Zoo-zoo DNA yuav muaj A260/A280 Qhov ratio ntawm 1.7-2.0.
Ib yam li ntawd, dab tsi yog qhov zoo DNA concentration? A zoo zoo DNA qauv yuav tsum muaj A260/A280 piv ntawm 1.7-2.0 thiab A260/A230 qhov piv ntawm ntau dua 1.5, tab sis txij li qhov rhiab heev ntawm cov txheej txheem sib txawv rau cov kab mob sib txawv, cov txiaj ntsig no tsuas yog yuav tsum tau coj los ua ib qho kev qhia rau kev coj dawb huv ntawm koj tus qauv.
Hais txog qhov no, NanoDrop xam DNA concentration li cas?
Koj muab tus nqi ntawm absorbance ntawm 260 nm los ntawm qhov tseem ceeb (nws yog 50 rau DNA ) thiab koj tau txais DNA concentration . Voila. (Ok nws yog technically A260 ntawm 10-mm tuab qauv uas tau muab faib ua 50; NanoDrop qhia A260 zoo li tus qauv yog 10mm tuab, zoo li hauv tus qauv cuvette).
Vim li cas peb thiaj li yuav tsum siv lub spectrophotometer los ntsuas peb DNA?
A spectrophotometer yog tuaj yeem txiav txim siab qhov nruab nrab ntawm cov nucleic acids DNA los yog RNA tam sim no nyob rau hauv ib tug sib tov, nrog rau lawv purity. Siv lub Beer-Lambert law nws yog muaj peev xwm txheeb xyuas tus nqi ntawm lub teeb absorbed rau qhov concentration ntawm absorbing molecule.
Pom zoo:
Yuav ua li cas koj pom qhov concentration ntawm DNA los ntawm absorbance?
DNA concentration yog kwv yees los ntawm kev ntsuas qhov nqus ntawm 260nm, kho qhov ntsuas A260 rau turbidity (tseem los ntawm absorbance ntawm 320nm), muab faib los ntawm cov khoom dilution, thiab siv kev sib raug zoo uas A260 ntawm 1.0 = 50µg / ml ntshiab dsDNA
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