RecoverEase protocol version D questions

Question

Hi Tech support team, We have received some questions regarding the new protocol of RecoverEase. Could you please tell us the answer? 
 In P5，step 25, &ldquo;Immediately spin the conical tube containing the dialysis tube for 5&ndash;6 seconds at 500&ndash;1000 &times; g. Do not spin longer than 6 seconds as this may cause the membrane to rupture&rdquo;. 
 Q1, Should we count 5-6 second when the centrifuge reach the appropriate speed which is 500-1000 x g, not from starting the spinning? I think usually it should be counted after reaching the desired speed, but I would like to confirm. 
 Q2, How immediately should we start spinning after dialysis? What could be the negative impact if we don&rsquo;t spin the tube immediately after dialysis? 
 
 Thank you for your support in advance. 
 Best regards, 
 Sayano Kinjo

agrafsky · Accepted Answer

Ordinarily, most centrifuges will begin application of the necessary force within less than a second of their starting, so it would be fine to start counting the 5-6 seconds as soon as the centrifugation begins. Obviously, if the particular centrifuge requires several seconds to reach the point where it is generating 500 x g, then it would be best to wait until it reaches the speed at which this force is generated. The main reason for centrifuging immediately is that the genomic DNA should not be allowed to become dehydrated and the conical tube cannot be sealed until the dialysis cap is replaced with the storage cap. Once the tube is sealed, then the gDNA may be stored.