E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)

Cat. No.: E1527100

QUALITÉ & PURETÉ BioReagent

Vue d’ensemble

Escherichia coli (E. coli) is a Gram-negative, short rod-shaped bacterium, measuring approximately 0.5 × 1–3 μm. It is motile by peritrichous flagella, does not form spores, and ferments various sugars to produce acid and gas. As a normal inhabitant of the intestinal tract of humans and animals, E. coli colonises the gut shortly after birth and remains throughout life, constituting nearly one‑third of the dry weight of faeces. The yield of DNA obtained from E. coli is high, and though the purity is generally moderate, it is sufficient for most molecular biology experiments.

CTAB (cetyltrimethylammonium bromide) is a cationic detergent. In solutions of high ionic strength, CTAB forms complexes with proteins and with most polysaccharides except acidic polyglycans, but it does not precipitate nucleic acids. Therefore, CTAB can be used to purify DNA from organisms that produce mucopolysaccharides, such as plants and certain Gram‑negative bacteria.

This product is a convenient kit for extracting genomic DNA from E. coli using the CTAB method. The extraction principle involves digestion of proteins by Proteinase K, precipitation of polysaccharides by CTAB, and subsequent heating to separate denatured proteins from DNA, followed by ethanol precipitation of the nucleic acid to obtain genomic DNA. The resulting DNA is suitable for downstream applications such as restriction enzyme digestion and PCR.

This reagent is for research use only and is not intended for clinical diagnosis or other applications.

Storage Unit No.	Storage Unit Name	Component No.	Component Name	Appearance	50T	Storage
E1527100NT	E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)(RT Components)	E1527100A	E. coli Lysis Buffer	Liquid	1.5ml	Room temperature
		E1527100B	CTAB Precipitation Solution	Liquid	10ml	Room temperature
		E1527100C	E. coli Wash Buffer	Liquid	100ml	Room temperature
		E1527100D	TE Buffer	Liquid	50ml	Room temperature
E1527100WT	E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)(2-8℃ Components)	E1527100E	Protein Removal Solution	Liquid	50ml	2-8℃.Store in the dark.
E1527100DT	E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)(-20℃ Components)	E1527100F	Proteinase K Solution	Liquid	0.5ml	-20℃

Note: Each component is individually packaged according to its storage temperature (Room temperature/2‑8 °C / -20 °C) upon shipment, with differentiated temperature‑controlled transportation to guarantee product performance. Orders containing materials requiring multiple temperature zones may be delivered in separate batches; please refer to the actual received goods. Upon receipt, store components separately at the temperatures specified in the component list. The shelf‑life is 12 months from the date of manufacture.

Reagents, consumables and Equipments not provided

1.Centrifuge tubes, centrifuge, shaker, incubator or water bath

2.70% Ethanol

Protocol (For Reference Only)

1.Transfer 1.5 ml of bacterial culture to an EP tube. Centrifuge at 10,000 rpm for 2 min, discard the supernatant. Add another 1.5 ml of bacterial culture and repeat the centrifugation step once more.

2.Add 30 µl of Eco Lysis Buffer and 3 µl of Proteinase K Solution. Mix thoroughly and incubate at 37°C for 1 h.

3.Add 180 µl of CTAB Precipitation Solution. Mix thoroughly and incubate at 65°C for 10 min.

4.Add 780 µl of Protein Removal Solution. Mix thoroughly and centrifuge at 12,000 rpm for 5–10 min.

5.Transfer the supernatant to a new EP tube. Add 2 volumes of Eco Wash Buffer, mix well, and place at -20°C for 30 min or overnight.

6.Centrifuge at 13,000 rpm for 10 min, discard the supernatant. Wash the pellet with 70% ethanol.

7.Centrifuge at 13,000 rpm for 10 min, discard the supernatant, and air-dry the DNA pellet at room temperature.

8.Dissolve the pellet in 100 µl (or an appropriate volume) of TE Buffer and store at -20°C. Note: A larger volume of TE Buffer will result in a lower DNA concentration.

Precautions

1. For optimal lysis and maximum yield, use freshly prepared phage or host bacterial cultures.

2. Eco Lysis Buffer may solidify at low temperatures. It can be redissolved by warming in a 40–50°C water bath.

3. For your safety and health, please wear a lab coat and disposable gloves during the procedure.

Spécifications

Synonymes

Escherichia coli genomic DNA extraction kit

Spécifications et pureté

BioReagent

Grade

BioReagent

Application

DNA Extraction

Sensibilité

Light Sensitive

Stockage et expédition

Conditions de stockage de stockage

Store at 2-8°C,Protected from light,Room temperature,Store at -20°C

Expédié en

Normal,Wet ice,Ice chest + Ice pads

Stabilité et stockage

Each component has a shelf life of 12 months under corresponding storage conditions.

Contents & Storage

Storage Unit No.	Storage Unit Name	Component No.	Component Name	Appearance	50T	Storage
E1527100NT	E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)(RT Components)	E1527100A	E. coli Lysis Buffer	Liquid	1.5ml	Room temperature
		E1527100B	CTAB Precipitation Solution	Liquid	10ml	Room temperature
		E1527100C	E. coli Wash Buffer	Liquid	100ml	Room temperature
		E1527100D	TE Buffer	Liquid	50ml	Room temperature
E1527100WT	E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)(2-8℃ Components)	E1527100E	Protein Removal Solution	Liquid	50ml	2-8℃.Store in the dark.
E1527100DT	E. coli Genomic DNA Extraction Kit (CTAB Precipitation Method)(-20℃ Components)	E1527100F	Proteinase K Solution	Liquid	0.5ml	-20℃

Note: Each component is individually packaged according to its storage temperature (Room temperature/2‑8 °C / -20 °C) upon shipment, with differentiated temperature‑controlled transportation to guarantee product performance. Orders containing materials requiring multiple temperature zones may be delivered in separate batches; please refer to the actual received goods. Upon receipt, store components separately at the temperatures specified in the component list. The shelf‑life is 12 months from the date of manufacture.