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.pdf
P2069
Phenol:Chloroform:Isoamyl Alcohol 25:24:1 Saturated with 10 mM Tris, pH 8.0, 1 mM EDTA
Supplied with Equilibration buffer, for molecular biology
Synonym(s):
DNA extraction
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100 ML
$119.00
400 ML
$363.00
$119.00
Estimated to ship onMarch 29, 2025
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100 ML
$119.00
400 ML
$363.00
About This Item
UNSPSC Code:
12190000
NACRES:
NA.52
grade:
for molecular biology
assay:
≥99.0%
application(s):
agriculture
$119.00
Estimated to ship onMarch 29, 2025
Recommended Products
grade
for molecular biology
Quality Level
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
For applications requiring a higher pH, such as the isolation of large intact genomic DNA, addition of the Equilibration Buffer is recommended.
Packaging
Supplied with P3803 and a separate bottle of equilibration buffer to adjust the phenol phase to pH 8.0 ± 0.2.
Suitability
This lot of phenol:chloroform:isoamyl alcohol 25:24:1 was use tested by extracting a crude preparation of pBR322 DNA preparation from E. coli. Based on evaluation by agarose gel electrophoresis the extracted DNA was not degraded or reduced in concentration. The same procedure was applied to purified pBR322 DNA and lambda Hind III digest DNA. Based on electrophoretic evaluation the purified preparations were not detectably degraded or reduced in concentration.
Preparation Note
Add the entire contents of the small bottle of Equilibration Buffer to the large bottle of Phenol:Chloroform:Isoamyl Alcohol. Mix gently and allow the phases to separate before use, approximately 2-4 hours. This will adjust the pH of the Phenol solution from pH 6.7 ± 0.2 to 8.0 ± 0.2.
Other Notes
After the addition of Equilibration Buffer, the product is stable for 6 months vs. 2 years without added buffer. Storage at -20 °C protected from light extends the shelf life to greater than 1 year after the addition of Equilibration Buffer.
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Product No.
Description
Pricing
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
Storage Class
6.1A - Combustible acute toxic Cat. 1 and 2 / very toxic hazardous materials
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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Protocols
Method for reverse transcription of RNA into DNA. Uses a premixed reagent that contains reverse transcriptase, dNTPs, primers, RNase inhibitor and buffer. Fast generation of cDNA.
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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-
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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.
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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.
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What is the Department of Transportation shipping information for this product?
1 answer-
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.
Helpful?
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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?
1 answer-
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?
1 answer-
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.
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