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What is Buffer AL and what does it do?
Buffer AL is used in DNA extraction and causes cell lysis to expose the DNA. Buffer AL is used during DNA isolation using QIAamp and DNeasy protocols. Buffer AL is stable for 1 year when stored closed at room temperature (15-25°C). Preparation of Buffer AL/E is as such: Volume of Buffer AL (ml) Volume of 96-100% ethanol (ml) Bottle size (ml) 33 35 100 108 114 250 162 171 500 216 228 500
How much of each ingredient do I need to make a 1mL lysis buffer solution?
You need to specify, lysis buffer for bacteria or eukaryotic cell. The most common buffer cocktail for eukaryotic cells is composed by an hypertonic solution and cell disrupters. To answer this question the proper amounts of the ingredients for 1 mL would be: 8.77 mg of NaCl (150 mM, the hypertonic component) 10 microliters NP-40 [can be replaced by 1 microliter (0.1%) of Triton X-100, a detergent] 6.06 mg of Tris in 100 microliters [Tris hydroxymethyl aminomethane, make 10 mL of a 100X solution (that is, 605.5 mg in 10 mL water, adjust to pH 8.0)] 890 microliters of distilled water. In some cases can be added some protease inhibitors such as PMSF, leupeptin, Aprotinin, etc. at concentrations of 1 microgram/mL. Store at 4oC for one month. Now, to make a lysis buffer for bacteria, the composition is different. For 1 mL mix: 6.06 mg of Tris in 100 microliters, as before 17.53 mg of NaCl (300 mM) 100 microliters of PMSF [Phenylmethylsulfonyl Fluoride, make 10X solution (1.742 mg in 10 mL of water)] 980 microliters of distilled water add aprotinin, leupeptin, and pepstatin at final concentration of 1 microgram/mL. Usually, to break the bacteria cell wall it is useful a lab blender. Disruption of both, eukaryotic and bacteria cells, must be done in cold conditions, usually on ice-water bath.
How do prepare 1X TE Buffer from 5X TE Buffer?
1 ml of 5X TE in 4ml distilled water (or).......if u want 100 ml just multiply 1 and 4 with 20....you will get 20 ml 5X TE in 80 ml distilled water
How do you change 100 to 3 significant figures?
Make it (100.) with a period at the end of the 100.
How do you make 0.3 percent Triton X-100?
triton x-100 stock percent? 100% ?
What is the function of tritonx100 in DNA extraction?
Triton X-100 is used as a lysis buffer for DNA separation.
Function of Triton X-100 in Lysis buffer?
It dissolves the cell mebrane. Keep in mind that some people do not use Triton X-100 in cell lysis, but instead sonificate... Also Triton X-100 can be replaced by a whole lot of other stuff (CHAPS, Igepal, etc) Hope to have givin you enough info
Role of tritonx100 in protein isolation?
TritonX-100 was used for Remove the SDS-From the crude protein, during homogenization the cell lysis buffer as contain SDS otherwise no need.
What is Buffer AL and what does it do?
Buffer AL is used in DNA extraction and causes cell lysis to expose the DNA. Buffer AL is used during DNA isolation using QIAamp and DNeasy protocols. Buffer AL is stable for 1 year when stored closed at room temperature (15-25°C). Preparation of Buffer AL/E is as such: Volume of Buffer AL (ml) Volume of 96-100% ethanol (ml) Bottle size (ml) 33 35 100 108 114 250 162 171 500 216 228 500
How much of each ingredient do I need to make a 1mL lysis buffer solution?
You need to specify, lysis buffer for bacteria or eukaryotic cell. The most common buffer cocktail for eukaryotic cells is composed by an hypertonic solution and cell disrupters. To answer this question the proper amounts of the ingredients for 1 mL would be: 8.77 mg of NaCl (150 mM, the hypertonic component) 10 microliters NP-40 [can be replaced by 1 microliter (0.1%) of Triton X-100, a detergent] 6.06 mg of Tris in 100 microliters [Tris hydroxymethyl aminomethane, make 10 mL of a 100X solution (that is, 605.5 mg in 10 mL water, adjust to pH 8.0)] 890 microliters of distilled water. In some cases can be added some protease inhibitors such as PMSF, leupeptin, Aprotinin, etc. at concentrations of 1 microgram/mL. Store at 4oC for one month. Now, to make a lysis buffer for bacteria, the composition is different. For 1 mL mix: 6.06 mg of Tris in 100 microliters, as before 17.53 mg of NaCl (300 mM) 100 microliters of PMSF [Phenylmethylsulfonyl Fluoride, make 10X solution (1.742 mg in 10 mL of water)] 980 microliters of distilled water add aprotinin, leupeptin, and pepstatin at final concentration of 1 microgram/mL. Usually, to break the bacteria cell wall it is useful a lab blender. Disruption of both, eukaryotic and bacteria cells, must be done in cold conditions, usually on ice-water bath.
How do you convert 1x buffer to 10x buffer?
The stocks are commonly labeled as X factors such as 10X, 5X, 100X etc. X-factor indicates that the solution is concentrated and must be diluted usually with water to 1X concentration for use. For eg: - A 100X concentrated solution should be diluted to 100 fold. to convert 1X to 10X take one ml of 1x buffer in a measuring cylinder and dilute it to make it 10 ml. its now 10x buffer.
In this procedure a 10x buffer is used if you have a 5x buffer and you want a total volume of 200ul what dilution would you perform to achieve the desired 1x concentration of buffer?
10x to 1x is a 1:10 dilution Therefore, add 1 part buffer, 9 parts DI-water If 100uL is 10uL (1 part buffer) and 90uL (9 parts DI-water) Then, 200ul (100 x 2) is 20uL (1 part buffer) and 180uL (9 parts DI-water)
How do you make 50 mM potassium phosphate buffer pH 7.4?
1 M Sodium Phosphate Buffer Stock Solution (1 liter) Protocol # Solution A: Dissolve 138.0 g NaH2PO4?H2O in 1 liter dH2O (pH 7.0). # Solution B: Dissolve 142.0 g Na2HPO4 in 1 liter dH2O (pH 7.0). # Mix 423 ml Solution A with 577 ml Solution B. # Autoclave and store at room temperature.
What does buffer aw1 and aw2 do?
Buffer AW1 contains Guanidinium Chloride (guanidine hydrochloride). This is used to denature proteins in your sample. They will then flow through the column and will be discarded with the wash. Buffer AW2 is essentially 70% EtOH. 30 mls of 100% EtOH is added to the 13 mls of "concentrate"included in the bottle. 70% EtOH is used to remove salts from your column and aid in purifying your DNA.
How do prepare 1X TE Buffer from 5X TE Buffer?
1 ml of 5X TE in 4ml distilled water (or).......if u want 100 ml just multiply 1 and 4 with 20....you will get 20 ml 5X TE in 80 ml distilled water
What numbers make 100?
any number A and 100-A would make 100
How many tens make up a thousand?
100 tens make up 1000
