4. Decant the supernatant and briefly vortex the pellet in the residual water. 5. In the following order, add 200μL Yeast Lysis Buffer (see below for recipe), 200μL phenol:chloroform:isoamyl alcohol (25:24:1), and 0.3 g of acid-washed glass beads (Sigma ). 6. Vortex for 3-4 m...
Preparation of Plasmid DNA by Alkaline Lysis with SDS: Maxipreparation. There are probably as many "miniprep" recipes as there are laboratories doing molecular biology. The following protocol is derived from the alkaline lysis recipe originally described by Birnboim and D J Sambrook,DW Russell - ...
However, in the preferred embodiment of the bulk leaf maceration method, the following recipe is used: 0.1% w/v Tween-80; 0.04 M Tris-Cl, pH 7.7; 0.15 M NaCl; 0.1% w/v BSA-Pentex fraction V; 0.01% w/v sodium azide; 200 mM EDTA. The color dye tartrazin may optionally be added...
25. We screened a series of BS conditions and identified a BS recipe (UBS-1), consisting of 10:1 (vol/vol) 70% and 50% ammonium bisulfite. We incubated a 5-mer DNA oligo AGCGA (Supplementary Table1) with UBS-1 at 98 °C, and matrix-assisted laser desorption ionization time-of-...
2. Prepare Lysozyme Digestion Buffer (see recipe on page 15). To ~200 μL Lysozyme Digestion Buffer/sample, add fresh Lysozyme to obtain a final Lysozyme concentration of 20 mg/mL. 3. Harvest up to 2 × 109 Gram positive cells by centrifugation. If you are using a frozen cell pellet,...
Immunoblotting analysis was performed following the procedure outlined in our previous study [34]. To perform immunoprecipitation, add the primary antibody to the cell lysate and rock overnight at 4 °C. Protein A beads were thoroughly washed with lysis buffer, and the supernatant was analyzed ...
在制作酶谱、测定序列、制备探针等实验中需要高纯度、高浓度的质粒DNA,为此需要大量提取质粒DNA。大量提取的质粒DNA一般需进一步纯化,常用柱层析法和氯化绝梯度离心法。实验材料细菌试剂、试剂盒STE酚 氯仿NaClPEG乙醇仪器、耗材离心管离心机抽干机实验步骤1、取培养至对数生长后期的含pBS质粒的细菌培养液250 ml,4 ℃...
If you would rather run gels at a lower voltage for any reason, gels can be run at 100 V for 30 min in 1x TAE electrophoresis buffer. Be sure to make 1x TAE at step 7 (same recipe as in step 1) if this is your preferred method. 1. Prepare 1x TAE gel buffer. 0.5 L of 1...
If labeled, it is important to maintain osmolality of the final solution (e.g., blood+buffer) to maintain high label efficiency. In certain embodiments, buffers used in methods of the invention are designed to prevent lysis of blood cells or any other cells, facilitate efficient binding of ...
Lysis buffer: (100 ml Recipe ) 10 mM Tris-HCl pH7.5 - 0.5 ml of 2M 10 mM EDTA- 2 ml of 0.5M 10 mM NaCl- 0.2 ml of 5M 0.5% (w/v)Sarkosyl- 0.5gm (N-Lauroylsarcosine,Sigma # L-9150) 1mg/ml Proteinase K- add fresh each time (stored in freezer). ...