How do you make 10mg ml Ethidium Bromide?

How do you make 10mg ml Ethidium Bromide?

Requirements

1. Step 1: Weigh out 100 mg ethidium bromide conical flask / beaker / 15-ml polypropylene centrifuge tube. Add 7 – 8 ml water.
2. Step 2: Mix until all ethidium bromide dissolves completely. This may take a long time.
3. Step 3: Adjust the volume to 10 ml with Deionized / Milli-Q water.

How much EtBr do I add?

(Optional) Add ethidium bromide (EtBr) to a final concentration of approximately 0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr binds to the DNA and allows you to visualize the DNA under ultraviolet (UV) light. CAUTION: EtBr is a known mutagen.

How do you dilute EtBr?

Dilute solutions containing EtBr to concentration <0.034% w/v (34mg/100mL). Add 10mL of fresh bleach for every 1mg EtBr. Stir at room temperature for at least 4 hours. The EtBr is converted to the physiologically inactive product 2-carboxybenzophenone.

What happens if you add too much Ethidium Bromide?

Adding too much ethidium on your gel can cause a lot of background fluorescence when visualising as well. Note that the SYBR Gold emission spectra is different from Ethidium Bromide as well so you might need a different filter on your imaging dock to see SYBR Gold-stained samples.

What is ethidium bromide used for?

Ethidium Bromide (EtBr) is sometimes added to running buffer during the separation of DNA fragments by agarose gel electrophoresis. It is used because upon binding of the molecule to the DNA and illumination with a UV light source, the DNA banding pattern can be visualized.

Is EtBr light sensitive?

The photo to the left shows a bottle of EtBr. It is always kept in light-sensitive bottles because of its photosensitivity, and concentrated stocks should be handled with caution. If the gel shows bands after EtBr staining, a polaroid photograph is taken as a permanent record.

How do you use EtBr?

Prepare enough 0.5µg/ml EtBr in water or buffer to completely submerge the gel. This solution is stable for 1-2 months at room temperature in the dark. After the run submerge the gel in the staining solution for 15-30 minutes (depending upon gel thickness).

How do you make a working solution of EtBr?

Ethidium Bromide Solution Preparation and Recipe

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 10 g of Ethidium bromide to the solution.
3. Stir on a magnetic stirrer for several hours to ensure that the dye has dissolved.

How much EtBr is toxic?

SYBRsafe was toxic at concentrations as low as 1 microgram/ml, whereas EthBr toxicity was not observed until 250micrograms/ml.

What could go wrong in gel electrophoresis?

Problems with the Gel, Current and Buffer The concentration of the gel must also be correct to avoid errors. If the concentration is too high or too low, the fragments will migrate either too slowly or too quickly. This will lead to errors in resolving the different bands.

Is ethidium bromide safe?

Although it is an effective tool, its hazardous properties require special safe handling and disposal procedures. EtBr is a potent mutagen (may cause genetic damage), and moderately toxic after an acute exposure. EtBr can be absorbed through skin, so it is important to avoid any direct contact with the chemical.

How bad is EtBr?

EtBr is a potent mutagen (can cause genetic damage), and moderately toxic after an acute exposure. EtBr can be absorbed through skin, so it is important to avoid any direct contact with the chemical. The powder form is considered an irritant to the upper respiratory tract, eyes, and skin.

What is the dosage of recofol in intensive care?

Induction & maintenance of general anesth. Sedation during intensive care. Induction of general anesth Adult <55 yr Initially 40 mg by slow IV bolus at intervals of 10 sec until onset of anesth. Usual dosage: 2-2.5 mg/kg. Childn >8 yr 2.5 mg/kg by slow IV until onset of anesth.

How much recofol is needed for general anesth?

Induction of general anesth Adult <55 yr Initially 40 mg by slow IV bolus at intervals of 10 sec until onset of anesth. Usual dosage: 2-2.5 mg/kg. Childn >8 yr 2.5 mg/kg by slow IV until onset of anesth. Maintenance of general anesth Adult 4-12 mg/kg/hr by continuous infusion. 25-50 mg by repeat bolus inj depending on response.

How big is a pack of recofol ampoules?

Recofol is a fast acting anesthetic for intravenous administration. sedative effect when performing diagnostic or surgical procedures during local or regional anesthesia. The release of the drug is produced in ampoules with a capacity of 20 ml. Inside a pack of 5 such ampoules.