Saures Phenol:Chloroform, pH 4,5 (mit IAA, 125:24:1)
Saures Phenol:Chloroform, pH 4,5 (mit IAA, 125:24:1)
Invitrogen™

Saures Phenol:Chloroform, pH 4,5 (mit IAA, 125:24:1)

Saures Phenol:Chloroform:IAA (125:24:1) ist vorgemischt und hat einen pH-Wert von 4,5 ± 0,2. Lieferung in einer Flasche mit 400 mlWeitere Informationen
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KatalognummerMenge
AM9720100 ml
AM9722400 mL
Katalognummer AM9720
Preis (EUR)
173,65
Online Exclusive
189,00
Ersparnis 15,35 (8%)
Each
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Menge:
100 ml
Recurring order eligible. Learn more »
Preis (EUR)
173,65
Online Exclusive
189,00
Ersparnis 15,35 (8%)
Each
Zum Warenkorb hinzufügen
Ask our AI about this Product
Saures Phenol:Chloroform:IAA (125:24:1) ist vorgemischt und hat einen pH-Wert von 4,5 ± 0,2. Lieferung in einer Flasche mit 400 ml Inhalt. Bei der RNA-Extraktion dient Phenol/Chloroform/IAA zur Entfernung von DNA (sie wird in der organischen Phase geteilt), trägt zur Stabilisierung der Interphase bei und verhindert die Schaumbildung beim Mischen. Die Vorbereitung von Phenol für Anwendungen in der Molekularbiologie ist wegen seiner toxischen und korrosiven Beschaffenheit ein zeitaufwändiges und oft gefährliches Verfahren. Vorgemischte, qualitätsgeprüfte, gesättigte Ambion® Phenole sind gebrauchsfertig und eliminieren so Probleme bei der Handhabung und bieten eine praktischere, sicherere und einfachere Alternative zur Vorbereitung von Lösungen aus kristallinem Phenol.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications
Chemischer Name oder MaterialPhenole
VerpackungsartFlasche
ProduktlinieAmbion™
ReinheitMolekularbiologische Qualität
Menge100 ml
VersandbedingungRaumtemperatur
FormFlüssig
pHpH 4,5
Unit SizeEach
Inhalt und Lagerung
Bei 4 °C oder -20 °C lagern.
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Häufig gestellte Fragen (FAQ)

Do you have any information on DNA and RNA purification using phenol chloroform and alcohol precipitation?

Phenol extraction of proteins:

Phenol extraction is frequently used to remove proteins from nucleic acid solutions. A common protocol is to add an equal volume of buffer-saturated phenol or phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) to an aqueous nucleic acid solution, vortex, and centrifuge at 14,000 x g for 1 min to separate the phases.

Studies at Thermo Fisher Scientific have shown that the concentration of NaCl in the aqueous solution should not exceed 0.5 M for good recovery of DNA. Residual phenol can be removed from the aqueous phase by extraction with an equal volume of chloroform or ether. After extraction, DNA is usually precipitated with ammonium acetate and ethanol as described in another protocol on this server. Ref. Karger, B. D. (1989) FOCUS 11, 14.

A good source of general information on the properties of phenol can be found in Wallace, Donald M. “Large and Small-Scale Phenol Extractions”. Methods in Enz. Volume 152 guide to Molecular Cloning Techniques. 1987. Academic Press, Inc. Berger and Kimmel, eds. Chap.4, pg 33-41.

(a) At pH 5 to 6 DNA is selectively retained in the organic phase and interphase, leaving RNA in the aqueous phase. Therefore a pH greater than 7 is needed if DNA is to be extracted.

(b) At pH values below 7.6, poly A+ RNA is lost to the organic phase if chloroform is not present.

(c) Optimal RNA yields in phenol extraction are obtained if the salt concentration is less than 0.15 M NaCl. Salt concentration in the sample is not a factor for larger DNA molecules.

To store RNA after extraction use DEPC-treated water.

What is the recommended protocol for phenol-extraction removal of proteins from nucleic acid containing solutions?

Below is a commonly used protocol:

(1) Add an equal volume of buffer-saturated phenol or phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) to an aqueous nucleic acid solution. Note: for RNA solutions, acid-phenol is recommended.

(2) Vortex, and centrifuge at 14,000 x g for 1 min to separate the phases.

(3) Residual phenol can be removed from the aqueous phase by extraction with an equal volume of chloroform or ether.

(4) After extraction, DNA/RNA is usually precipitated with ammonium acetate and ethanol.

What is the optimal pH of the phenol:chloroform mixture for isolation of DNA?

Partitioning of the nucleic acids in phenol is pH dependent. At pH 7.0 or higher, both DNA and RNA partition into the aqueous phase. At an acidic pH (below 7.0) DNA is denatured and will move into the organic phase, but the RNA remains in the aqueous phase. The mixture should be adjusted to at least pH 7.4 for work with DNA.

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