The contents of the Tris base are added to the phenol:chloroform:isoamyl alcohol mixture, and the contents are well mixed. The amount added is 6.5 mL equilibration buffer per 100 mL phenol, designed so that only one treatment is necessary to raise the pH. Allow the mixture to separate, which may take 2-4 hours. The upper aqueous phase is removed and discarded. The remaining lower organic phase is divided into aliquots and stored at -20 °C. The solution should be stable for up to 12 months. The pH of the phenol should be raised from 6.7 to about 7.9-8. Only one treatment is needed.
Fortfahren mit
P2069
Phenol:Chloroform:Isoamylalkohol 25:24:1, gesättigt mit 100 mM TRIS pH 8,0; 1 mM EDTA
Supplied with Equilibration buffer, Molecular Biology
Synonym(e):
DNA extraction
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| Ihnen/SKU | Verfügbarkeit | Preis |
|---|---|---|
100 mL | Warenkorb auf Verfügbarkeit prüfen | CHF 251.00 |
400 mL | Warenkorb auf Verfügbarkeit prüfen | CHF 717.00 |
Über diesen Artikel
NACRES:
NA.52
UNSPSC Code:
12190000
Assay:
≥99.0%
Grade:
Molecular Biology
CHF 251.00
Warenkorb auf Verfügbarkeit prüfen
Technischer Dienst
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Unterstützung erhaltengrade
Molecular Biology
Quality Segment
assay
≥99.0%
form
liquid
pH
6.5-6.9(pH of phenolic phase), 7.8-8.2(pH of phenolic phase after the addition of Equilibration Buffer)
application(s)
agriculture
storage temp.
2-8°C
Application
Die Zugabe des Äquilibrierungspuffers wird für Anwendungen empfohlen, für die ein höherer pH-Wert erforderlich ist, wie beispielsweise die Isolierung von großer intakter genomischer DNA.
Diese Charge von Phenol:Chloroform:Isoamylalkohol 25:24:1 wurde durch die Extraktion eines Rohpräparats des pBR322-DNA-Präparats aus E. coli geprüft. Die Beurteilung mittels Agarose-Gelelektrophorese ergab, dass die extrahierte DNA nicht abgebaut oder in ihrer Konzentration verringert worden war. Dasselbe Verfahren wurde bei gereinigter pBR322-DNA und Lambda Hind III Digest DNA angewendet. Die Beurteilung mittels Elektrophorese ergab, dass die gereinigten Präparate nicht nachweisbar abgebaut oder in ihrer Konzentration verringert worden waren.
Packaging
Im Lieferumfang sind P3803 und eine separate Flasche mit Äquilibrierungspuffer enthalten, um die Phenolphase auf einen pH-Wert von 8,0 ± 0,2 einzustellen.
Preparation Note
Den gesamten Inhalt der kleinen Flasche mit Äquilibrierungspuffer in die große Flasche mit Phenol:Chloroform:Isoamylalkohol geben. Vorsichtig mischen und vor Gebrauch ca. 2–4 Stunden warten, bis sich die Phasen getrennt haben. Dadurch wird der pH-Wert der Phenollösung von 6,7 ± 0,2 auf 8,0 ± 0,2 eingestellt.
Other Notes
Nach der Zugabe von Äquilibrierungspuffer ist das Produkt 6 Monate haltbar, ohne Zugabe von Puffer ist es 2 Jahre haltbar. Durch lichtgeschützte Lagerung bei -20 °C kann die Haltbarkeit nach der Zugabe des Äquilibrierungspuffers auf mehr als 1 Jahr verlängert werden.
1 of 1
Dieser Artikel | |||
|---|---|---|---|
| assay ≥99.0% | assay ≥99.0% | assay ≥99.0% | assay - |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| grade Molecular Biology | grade Molecular Biology | grade Molecular Biology | grade Molecular Biology |
| application(s) agriculture | application(s) agriculture | application(s) life science and biopharma | application(s) - |
| form liquid | form liquid | form liquid | form liquid |
| pH 6.5-6.9(pH of phenolic phase), 7.8-8.2(pH of phenolic phase after the addition of Equilibration Buffer) | pH 6.5-6.9(pH of phenolic phase) | pH 4.3-4.7 | pH 4.1-4.5 (25 °C) |
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signalword
Danger
Hazard Classifications
Acute Tox. 3 Oral - Acute Tox. 4 Dermal - Acute Tox. 4 Inhalation - Aquatic Chronic 2 - Carc. 2 - Eye Dam. 1 - Muta. 2 - Repr. 2 - Skin Corr. 1B - STOT RE 1 Oral - STOT RE 2 - STOT SE 3
target_organs
Central nervous system, Liver,Kidney, Nervous system,Kidney,Liver,Skin
Lagerklasse
6.1A - Combustible acute toxic Cat. 1 and 2 / very toxic hazardous materials
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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Hello, I used P2069 for DNA extraction from mouse tails. After adding the equilibration buffer, I wait for 2~4hrs until phase seperation. At this point, I have a question. I heard that lower phase is the phase I utilize, so Can I aliquot the lower phase?
1 answer-
Helpful?
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Hello, I used this material for DNA extraction from mouse tail. After mixing the equilibration buffer, there was two phase. Which phase should I use? Also, Can I aliquot the liquid?
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The lower phase should be used for DNA extraction. It is possible to aliquot the liquid immediately after mixing, before the phases separate.
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After equilibrating with the supplied Tris buffer, can I aliquot this for use, if so, should I just top each aliquot with additional Tris pH 8?
1 answer-
Please see the link below to review the product datasheet. Note that the prepration instructions require the addition of the entire vial (6.5 mL) of equilabration buffer to the contents of the product. Aliquot immediately after the gentle mixing step. During the 2 - 4 hour incubation period the mixure will phase separate. At this point the aliquots may be stored for long term use according to the instructions.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/144/892/p2069dat.pdfHelpful?
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There is an Equilibration Buffer (Product B5658) that is included with Product P2069, Phenol:Chloroform:Isoamyl Alcohol 25:24:1. Is B5658 also available separately?
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No, it is only available as part of P2069.
Helpful?
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I see that you list two equilibrated Phenol:Chloroform:Isoamyl alcohol products. When should I use Product P3803, and when should I use Product P2069?
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In short, P3803 is for RNA work, and P2069 is for DNA work. Product P2069 consists of one bottle of Product P3803 and one bottle of Product B5658. The B5658 is added to the P3803 to raise the pH of the organic phase from 6.7 to 8.0; pH 8.0 is not suitable for RNA work, because RNA can degrade at the higher pH.
Helpful?
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I ordered Product P2069, Phenol:Chloroform:Isoamyl Alcohol 25:24:1; I got one bottle of the Phenol:Chloroform:Isoamyl Alcohol and one bottle of B5658, Equilibration buffer. When and how am I supposed to use the B5658?
1 answer-
B5658 is a Tris buffer that is used to raise the pH of the phenol:chloroform:isoamyl alcohol mixture, to make it suitable for isolation of genomic DNA. The contents of the two bottles need to be well mixed. Then allow the mixture to separate; this may take 2-4 hours. This procedure should raise the pH of the phenol phase from 6.7 to about 8.0. Only one "treatment" is needed. This solution should be stable for up to 12 months.
Helpful?
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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There are two layers in the bottle of Product P2069, Phenol:Chloroform:Isoamyl Alcohol 25:24:1; which layer should I use?
1 answer-
The lower phase is the phase that is to be used.
Helpful?
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