If you are looking for lysis buffer with ratio buffer:sample 1:4/1:2 then detergents or ferments seems to be obligatory. Depending of your needs it could be good or not..

We use an Ammonium Chloride Lysis solution (0.8% NH4Cl, 0.1 mM EDTA in water buffered with KHCO3 to achieve a final pH of 7.2 - 7.6), the recipe originates from StemCell Technologies.  1 part blood + 9 parts Lysis solution, on ice 5 minutes, a gentle vortex, then on ice another 5 minutes with a second gentle vortex.  Spin down white blood cells.  You can adapt your solution concentration so that it fits into your required ratio.

Helo Elizabeth

Thank You for the protocol.

I am seeking any other methods where the amount of buffer to be added : to sample is 2:1/4:1 instead of 9:1 or above as in most of the protocols.

Let us assume we have prepared 10 X concentration of the lysis buffer, the working conc is 1X. but to decrease the volume of the buffer we use, have u tried using 5X-8X as working concentration and checked if u might need lesser buffer to sample ratio and does that really work in lysis

Hey Daniil

What did u meant by ferments.?

and if detergents? can u specify any protocols

Like almost all the available commercial kit methods have buffer: sample ranging from 9:1 to 15:1

I wish to know if there are any other protocols where the same ratio is 2:1-4:1 at maximum.

The quality of lysis will depends of many factors. You have not specified nor materials nor your goal on the next step. Impurities, components and shadows of RBCs could be good or not for you.

Hello Danil

My goal is to seek any protocol which uses less volume of lysis buffer for more sample (1:2 1:1 in ratio) so as to lyse the RBC in my blood sample.

All the available methods use a lot of 1X lysis buffer for very less sample volume. I was wondering if there are any methods where the lysis buffer to be added is minimal and the process of lysis is faster.

Ultimate aim is to isolate DNA from whole blood.

Since this is for DNA isolation, my approach will work.  We use the 10X above and pellet the WBC to extract the DNA.  Is there any particular reason why you require the low ratio? You will still have to pellet the WBC before beginning the DNA isolation because the high hemoglobin concentration will cause problems. We have never tried a more concentrated stock at a reduced ratio. That would imply considering the isotonicity of the blood in the calculation of the more concentrated stock.  You can also consider the Paxgene kits by Qiagen.  They have a kit for both human and mouse blood.

we have use sucrose solution for RBC lysis in dna isolation from whole blood (109 gram of sucrose dissolve in 1000ml +add 5ml 1 Molar MgCl2  and 10 ml titron x-100) mix the solution than centrifuge it ar 7000rpm for 7 mins.U get best result.

Hello Preeti 

Can you share the entire protocol u have followed. thank you for the information.