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.
P2069
Phenol:Chloroform:Isoamyl Alcohol 25:24:1
Supplied with Equilibration buffer, Molecular Biology
Synonyma:
DNA extraction
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Vybrat velikost
Změnit zobrazení
| Velikost balení | Skladová položka | Dostupnost | Cena |
|---|---|---|---|
| 100 mL | Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti. | 2 970,00 Kč | |
| 400 mL | Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti. | 9 000,00 Kč |
O této položce
NACRES:
NA.52
UNSPSC Code:
12190000
Assay:
≥99.0%
Grade:
Molecular Biology
2 970,00 Kč
Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti.
Technický servis
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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
Phenol:Chloroform:Isoamyl Alcohol has been used in the preparation of RNA for the study of total RNA isolation from myocardial biopsies.[1] It has been also used to extract DNA during Human Androgen Receptor (HUMARA) Clonality Assay.[2]
For applications requiring a higher pH, such as the isolation of large intact genomic DNA, addition of the Equilibration Buffer is recommended.
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.
Packaging
Supplied with P3803 and a separate bottle of equilibration buffer to adjust the phenol phase to pH 8.0 ± 0.2.
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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Tato položka | |||
|---|---|---|---|
| assay ≥99.0% | assay ≥99.0% | assay ≥99.0% | assay - |
| 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) - |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| 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
Skladovací třída
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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67-66-3
CAS No.
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Protokoly
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.
Související obsah
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Bacterial flagellin is an important antigen in inflammatory bowel disease, but the role of flagellin-specific CD4+ T cells in disease pathogenesis remains unclear. Also unknown is how changes in intestinal microbiome intersect with those in microbiota-specific CD4+ T cells. We
Jacob F Warner et al.
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The sea anemone Nematostella vectensis is an emerging research model to study embryonic development and regeneration at the molecular and global transcriptomic level. Transcriptomics analysis is now routinely used to detect differential expression at the genome level. Here we present
Globální číslo obchodní položky
| Skladová položka | GTIN |
|---|---|
| P2069-100ML | 04061837898709 |
| P2069-400ML | 04061837898723 |
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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.
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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.
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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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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?
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The lower phase is the phase that is to be used.
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