Home About Us News Products Technical Support Download Selling Network Recruitment Feedback Contact Us
Product class
DNA Markers(Ladders)
PCR and RT-PCR
Plasmid Isolation
PCR Product/Gel Purification
Genomic DNA Isolation
RNA Isolation
Biochemicals
Protein Analysis
Electrophoresis Products
Molecular Biology Reagent
DNA/RNA Spin column
Lab Instruments
Products
Technical Support You are here:Home >> Technical Support
Preparation of laboratory reagents and buffers

 1M Tris-HCl (pH7.4, 7.6, 8.0)
Component concentration: 1 M Tris-HCl
Preparation of volume: 1 L
Preparation method:
1. Weighing 121.1 g Tris placed in 1 L beaker.
2. Add about 800 ml of deionized water, stir to dissolve.
3. The following table by adding concentrated hydrochloric acid to adjust the amount of required pH.
4. The solution to volume 1 L.
5. Autoclave, the room temperature.
Note: should the solution to cool to room temperature before it is set pH, Tris, pH values because of temperature differences in a large, temperature rises 1 ℃, the solution pH value of about 0.03 units lower.
  
10×TE Buffer (pH7.4, 7.6, 8.0)
Component concentration: 100 mM Tris-HCl, 10 mM EDTA
Preparation of volume: 1 L
Preparation method:
1. Capacity to take the following solution, placed in 1 L beaker.
2. To the beaker, add about 800 ml of deionized water and mix.
3. The solution to volume 1 L, the high-temperature autoclave.
4. Room temperature.
  
1.5M Tris-HCl (pH8.8)
Component concentration: 1.5 M Tris-HCl
Preparation of volume: 1 L
Preparation method: 
1. Weighing 181.7 g Tris placed in 1 L beaker. 
2. Add about 800 ml of deionized water, stir to dissolve. 
3. With concentrated hydrochloric acid to adjust pH value of 8.8. 
4. The solution to volume 1 L. 
5. Autoclave, the room temperature. 
Note: should the solution to cool to room temperature before it is set pH, Tris, pH values because of temperature differences in a large, high-temperature per 1 ℃, the solution pH value of about 0.03 units lower.
  
3M sodium acetate (pH5.2)
Component concentration: 3M sodium acetate
Preparation of volume: 100ml
Preparation method:
1. Weighing 40.8g NaAc·3H2O placed in 100-200ml beaker, add 40ml of deionized water on stirring dissolved
2. Acetic acid to adjust pH value of 5.2
3. Deionized water added to the solution volume to 100ml
4 autoclave, the room temperature.
  
PBS Buffer
Component concentration: 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4
Preparation of volume: 1 L
Preparation method:
1. Weighing the following reagent, placed in 1 L beaker.
2. To the beaker, add about 800 ml of deionized water, stir to dissolve.
3. Dropwise concentrated hydrochloric acid to adjust pH value to 7.4, then deionized water, the solution volume to 1 L.
4. Autoclave, the room temperature.
Note: The above goes for divalent cations in PBS Buffer, if necessary, in the formula to add 1 mM CaCl2 and 0.5 mM MgCl2.
  
10M  Ammonium acetate
Component concentration: 10 M ammonium acetate
Preparation of content: 100 ml
Preparation method: 
1. Weighing 77.1 g ammonium acetate, placed in 100 ~ 200 ml beaker, add about 30 ml of deionized water and stir to dissolve. 
2. Deionized water added to the solution volume to 100 ml. 
3. Using 0.22 mm membrane filter sterilization. 
4. Sealed bottle at room temperature. 
 
Note: The decomposition of ammonium acetate heat easily, it can not autoclave.
  
Tris-HCl balance of phenol
Preparation method:
1. Use of raw materials: Most of the commercially available colorless liquefied phenol is brighter without re-distillation can be used in molecular biology experiments. However, some liquefied phenol pink or yellow should be avoided. Also avoid using crystalline phenol, phenol crystallized at 160 ℃ for them to be re-distilled to remove oxidation products such as quinones, these oxidation products can cause the breaking phosphodiester bonds or cause cross-linking of RNA and DNA and so on. Therefore, the quality of phenol on DNA, RNA extraction is very important, and we recommend using high-quality molecular biology phenol. 
2. Operation Note: Phenol is extremely corrosive and can cause severe burns, the operation should wear gloves and protective glasses and so on. All operations should be carried out in the hood, and parts of phenol skin contact wash with plenty of water and wash with soap and water, avoid using alcohol. 
3. Phenol balance: because in the acidic pH, DNA distribution in the organic phase, the use of phenol phenol must be balanced before the pH value of 7.8 to more than balance the action of phenol is as follows:
  
① liquefied phenol should be stored at -20 ℃, at this time of phenol was crystallized state. Remove from the freezer of phenol at room temperature first to reach the room temperature, then phenol in 68 ℃ water bath to fully melt manipulation.
  
② adding hydroxyquinoline (8-Quinolinol) to a final concentration of 0.1%. The compound is a reducing agent, RNA enzyme inhibitors and metal ions are not completely weak chelating agent, also because of its yellow to help facilitate easy identification of the organic phase.
  
③ equal volume of 1 M Tris-HCl (pH8.0), using magnetic stirrer for 15 minutes, standing to make it fully stratified, remove the upper aqueous phase.
  
Repeat steps ④ ③.
  
⑤ adding an equal volume of 0.1 M Tris-HCl (pH8.0), using magnetic stirrer for 15 minutes, standing to make it fully stratified, remove the upper aqueous phase.
  
Repeat steps ⑥ ⑤, little remains of the upper aqueous phase.
  
Confirmed using pH strips ⑦ organic phase of the pH value is greater than 7.8.
  
⑧ will be placed in brown glass bottles phenol 4 ℃ stored.
  
Phenol/chloroform/isoamyl alcohol (25: 24: 1)
Preparation method:
1. Note: to remove the protein from the DNA samples are often used when the phenol / chloroform / isoamyl alcohol (25: 24: 1.) Protein denaturation and help make chloroform phase and organic phase separation, and isoamyl alcohol extraction process will help eliminate air bubbles appear.
2. Preparation Methods: Tris-HCl balance with an equal volume of phenol chloroform / isoamyl alcohol (24: 1) after mixing, into brown glass bottles at 4 ℃.
  
10%(W/V) SDS
Component concentration: 10% (W / V) SDS
Preparation of volume: 100ml
Preparation method:
1. Weighing 10g placed in high-purity SDS 100-200ml beaker, add about 80ml of deionized water, 68 ℃ adding dissolved
2. Dropwise concentrated hydrochloric acid to adjust pH value of 7.2
3. The solution to volume 100ml, the room temperature.
  
2N NaOH
Component concentration: 2 N NaOH
Preparation of content: 100 ml
Preparation method:
1. Capacity to take 80 ml of deionized water placed in 100 ~ 200 ml plastic beaker (NaOH large exothermic process of dissolution, it may burst the glass beaker).
2. Weigh 8 g NaOH carefully and gradually added to the beaker, while stirring add.
3. Until NaOH is completely dissolved, the volume of deionized water, the solution volume to 100 ml.
4. The solution into a plastic container, the room temperature.
  
2.5N HCl
Component concentration: 2.5N HCl
Preparation of content: 100 ml
Preparation method:
1. In 78.4 ml of deionized water by adding 21.6 ml of concentrated hydrochloric acid (11.6 N), uniformly mixed.
2. Room temperature.
  
5M NaCl
Component concentration: 5 M NaCl
Preparation of volume: 1 L
Preparation method:
1. Weigh 292.2 g NaCl placed in 1 L beaker, add about 800 ml of deionized water and mix to dissolve.
2. Deionized water added to the solution volume to 1 L, the amount is divided into small portions.
3. After autoclave, 4 ℃ preservation.
  
20%(W/V) Glucose
Component concentration: 20% (W / V) Glucose
Preparation of content: 100 ml
Preparation method:
1. Weigh 20 g Glucose placed in 100 ~ 200 ml beaker, add about 80 ml of deionized water, stirring to dissolve.
2. Deionized water added to the solution volume to 100 ml.
3. After autoclave, 4 ℃ preservation.
  
Solution I(plasmid extraction only)
Component concentrations: 25 mM Tris-HCl (pH8.0), 10 mM EDTA, 50 mM Glucose
Preparation of volume: 1 L
Preparation method:
1. Capacity to take the following solution, placed in 1 L beaker. 
2. After autoclave, 4 ℃ preservation.
3. Before use 50 ml of Solution I of each added to 2 ml of RNase A (20 mg / ml).
  
Solution II (plasmid extraction only)
Component concentration: 200mM NaOH, 1% (W / V) SDS
Preparation of volume: 500ml
Preparation method:
1. Capacity to take the following solution, placed in 500ml beaker
    
10% SDS       50ml
    
2N NaOH       50ml
2. Plus sterile water to volume 500ml, mix well
3. Room temperature, this solution save time, preferably not more than one month
Note: SDS is easy to produce bubbles, not stirred
  
Solution III (plasmid extraction only)
Component concentration: 3 M KOAc, 5 M CH3COOH
Preparation of content: 500 ml
Preparation method:
1. Weighing the following reagent, placed in 500 ml beaker.
2. Add 300 ml of deionized water and mix to dissolve.
3. Deionized water added to the solution volume to 500 ml.
4. After autoclave, 4 ℃ preservation.
  
0.5M  EDTA (pH8.0)
Component concentration: 0.5 M EDTA
Preparation of volume: 1 L
Preparation method:
1. Weigh 186.1 g Na2EDTA · 2H2O, placed in 1 L beaker.
2. Add about 800 ml of deionized water, stir.
3. With NaOH to adjust the pH value of 8.0 (about 20 g NaOH).
Note: pH value to 8.0, EDTA can be completely dissolved.
4. Deionized water added to solution to volume 1 L.
5. Amount into smaller portions, the high-temperature autoclave.
6. Room temperature.
  
1M  DTT
Component concentration: 1 M DTT
Preparation of content: 20 ml
Preparation method:
1. Weigh 3.09 g DTT, added to 50 ml plastic centrifuge tube.
2. Add 20 ml of 0.01 M NaOAc (pH5.2), dissolved in sterile filtered using 0.22 mm filter.
3. After the appropriate amount into smaller portions, -20 ℃ preservation.
  
10mM  ATP
Component concentrations: 10 mM ATP
Preparation of content: 20 ml
Preparation method:
1. Weigh 121 mg Na2ATP · 3H2O, added to 50 ml plastic centrifuge tube.
2. Add 20 ml of 25 mM Tris-HCl (pH8.0), stirring to dissolve.
3. After the appropriate amount into smaller portions, -20 ℃ preservation.

Home About Us News Products Technical Support Download Selling Network Recruitment Feedback Contact Us
Guangzhou Geneshun Biotech Ltd 粤ICP备08201538号
Tel:020-23312160 Fax:020-87056567
Email:geneshunbio@126.com