Nucleic Acid Extraction-Phenol Chloroform

Molecular Biology

nucleic acid extraction

Phenol-Chloroform

Extraction with phenol and phenol/chloroform mixtures is a universal method for purification of DNA and RNA. Proteins and restriction enzymes are removed by phenol and chloroform in disrupting protein secondary structure causing proteins to denature and precipitate from solution. Although each of these solvents is capable of performing this function alone, the two materials together remove proteins from solution much more effectively. Nucleic acids are recovered in the liquid phase.

Phenol and chloroform provided by Uptima are biotechnology grade products in order to offer you a safer and easier alternative to the use of crystalline phenol.

During phenol extraction, the precipitated proteins collect at an interphase between the aqueous and organic layers. It is very important to understand the effect of pH on the performance of phenol.

For the purification of DNA, the use of phenol at pH above 7.0 is desirable in order to collect DNA in the upper aqueous layer of a phenol extraction (phenol saturated, pH8: UP87336). At lower pH (4.5), DNA and proteins are collected at the interphase although RNA can be collected in the aqueous phase (phenol saturated, pH4,5: UP893305).

Phenol and chloroform are supplied as solution alone, premixed with chloroform and isoamyl alcohol to reduce the extraction time.

DNA purification:

Phenol saturated, pH8
Chloroform
Chloroform: isoamyl
Phenol: Chloroform
Guanidine Hydrochloride

RNA purification

Phenol saturated, pH4.5
Guanidine Hydrochloride

Phenol saturated, pH8	UP87336A	100ml	Chloroform	UP046904	950ml
	UP87336B	500ml
DNAse, RNAse, Protease : free Purity : 99,9% Technical Sheet			Stabilized with amylene DNAse, RNAse, Protease : free Purity : 99,9%
Chloroform : isoamyl	UP899254	450ml	Phenol: Chloroform	UP873357	100ml
alcohol 24 : 1 - Molecular Biology Grade	UP899255	950ml	isoamyl alcohol 25 : 24 : 1 - Molecular Biology Grade	UP873358	500ml
DNAse, RNAse, Protease : free Purity (chloroform and isoamyl alcohol) : 99,9%			Ready-to-use solution No manipulation of hazardous products Reduces extraction time Purity (phenol): 99,9% Security Sheet
Phenol saturated, pH 4.5	UP893305	100ml	Guanidine Hydrochloride	UP018380	500ml
Molecular Biology Grade	UP893306	400ml	Molecular Biology Grade
DNAse, RNAse, Protease : free Purity: 99,9% Technical sheet			Inhibition of RNAse and Protease Activity CH₅N₃HCl MW 95.53 CAS [50-01-1]

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