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Video: Yuav ua li cas koj pom qhov concentration ntawm DNA los ntawm absorbance?
2024 Tus sau: Miles Stephen | [email protected]. Kawg hloov kho: 2023-12-15 23:36
DNA concentration yog kwv yees los ntawm kev ntsuas nqus tau ntawm 260nm, kho qhov A260 ntsuas rau turbidity (ntsuas los ntawm nqus tau ntawm 320nm), muab faib los ntawm qhov sib txawv, thiab siv kev sib raug zoo uas tus A260 ntawm 1.0 = 50µg/ml ntshiab dsDNA.
Ib yam li ntawd, tib neeg nug, koj pom qhov concentration ntawm DNA li cas?
Txhawm rau txiav txim siab qhov concentration ntawm DNA hauv cov qauv qub, ua raws li hauv qab no:
- dsDNA concentration = 50 μg / mL × OD260 × dilution factor.
- dsDNA concentration = 50 μg / mL × 0.65 × 50.
- dsDNA concentration = 1.63 mg/mL.
Tsis tas 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.
Ua li no, puas DNA nqus ntawm 280 nm?
Qhov ratio ntawm absorbance ntawm 260 nm vs 280nm ua feem ntau siv los ntsuas DNA Kev sib kis ntawm cov kev daws teeb meem ntawm cov protein, txij li cov proteins (tshwj xeeb, cov amino acids aromatic) nqus teeb ntawm 280nm ua.
Koj siv NanoDrop li cas rau DNA concentration?
Yeej qhov nanodrop muab kev xaiv rau koj xaiv DNA , RNA, Proteins. Koj yuav tsum xaiv DNA , ces tso 2 ΜL ntawm dej (mili Q nyiam) xaiv "Blank" tom qab ntawd tso lwm 2 ΜL dej kom paub meej tias qhov ntsuas yog 0. Ces tso 2 ΜL ntawm koj tus qauv. Koj yuav tau txais qhov ntsuas.
Pom zoo:
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