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Q1 – The instructions for LPS say that 5 ml of bacteria is the maximum volume that gives the optimum purity. What about if we increase the bacterial volume as well as the reagents. For instance, start with 30 ml of culture ( 6 x 5ml culture) and use the reagents 6x the recommended volume. Will the purity be still optimum?
You can increase the starting volume. When doing this, you should increase the related reagents proportionally. We also recommend you to set the centrifuge over 10,000g in this case.
Q2 – What should the pellet look like after spinning down the sample in step 6?
The pellet should be opaque white and clearly visible. If you do not see a pellet, then we suggest increasing the amount of starting material (at least 2-fold to 10-fold).
Q3 – What is the yield of LPS using the kit?
The LPS kit has been reported to yield 30 ug of LPS from a 2ml culture of E. coli (OD600=1.0)