Customer Support - BAC DNA

Prepare a solution of 0.2N NaOH; 1% (w/v) SDS and substitute this for the current lysis buffer and follow the protocol as normal.

Because Cosmids are under 80kb (actually under 60kb) they don't require the modified BAC Lysis buffer in our kit. You could still use it; however, we've found standard lysis buffer improves the yield.

Be sure that the lysate was diluted as instructed in Step 7. The PSI Pressure Tip (included in the bag of columns) can be used to initiate or assist flow. Place the Pressure Tip on the end of a 3 mL Luer Lock syringe. Pull back the syringe plunger and place the tip on the top of the column. Depress the plunger to start flow. If flow doesn't start reseat the tip on the column. Maintain a flow rate of 1-2 drops/second until all of the lysate has been expelled from the column.

Do not release the pressure on the plunger while the syringe is seated on the column; If released the resulting vacuum may disrupt the column bed.

This procedure may also be needed to assist flow during the wash steps.

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