dna回收技术总结（DNA recovery technology summary）.docVIP

dna回收技术总结（DNA recovery technology summary） Rubber recovery I. measures for increasing the amount of reclaimed rubber: 1) increase the sample size at the time of electrophoresis. 2) the electrophoretic buffer was freshly prepared. 3) cut the glue as much as possible only with a strip of plastic, cut the volume of plastic: contains little pieces of purpose, glue will not have to, or else affect the recovery rate. 4) melt the cut of two or more pieces of glue, no matter how large, with a tube to the same column. 5) the solution added to sol can be a little more, so it is more conducive to the combination of DNA and membrane, but generally do not excess 750ul. 6) the key is through the plastic recycling alkali salt concentration, column solution (charge) and hydrophobic DNA and post combination, therefore, if the buffer PH is high, 10ul is added in the sol solution (PH 5, 3mol/L NaAC); in order to make a better interception in the membrane DNA molecule that can add 30% isopropanol in heated and dissolved after gel liquid. 7) place the column at room temperature for a few minutes (about 10 minutes) before the eluent is added, so that the alcohol is sufficiently volatile. 8) finally add some little eluent, minimize the volume recovered, with the general 30-50 l eluting (not too little, not wet film but not conducive to the drop in the eluent eluting); membrane central, in order to fully eluted with in membrane DNA. 9) after adding eluent, you can wash it in 55 degree water bath for 5 minutes, or put it in 50 degree water bath for more than 10 minutes, then wash it, or seal it for 4 degrees overnight, and then recycle it for second days. The result is good. 10) the eluent after centrifugation is added back to the adsorbent column and centrifuged again. Two. Detailed methods and steps for the recovery of several PCR products 1) common glue recovery If you want to glue recovery, should better use the kit, convenient, recovery rate is slightly higher, you really have to manua