Green Michael R, Sambrook Joseph
Cold Spring Harb Protoc. 2019 Feb 1;2019(2):2019/2/pdb.prot100479. doi: 10.1101/pdb.prot100479.
The standard method to recover fragments of DNA from polyacrylamide gels is the "crush and soak" technique. The eluted DNA is generally free of contaminants that inhibit enzymes or that are toxic to transfected or microinjected cells. The method requires time but little labor and results in recovery of <30%-90%, depending on the size of the DNA fragment. It can be used to isolate both double-stranded and single-stranded DNAs from neutral and denaturing polyacrylamide gels, respectively. The method is widely used to isolate synthetic oligonucleotides from denaturing polyacrylamide gels. DNA recovered from polyacrylamide gels by crushing and soaking is generally suitable for use as a hybridization probe, as a polymerase chain reaction (PCR) primer, and as a substrate in enzyme-catalyzed reactions.
从聚丙烯酰胺凝胶中回收DNA片段的标准方法是“挤压浸泡”技术。洗脱的DNA通常不含抑制酶活性或对转染细胞或显微注射细胞有毒的污染物。该方法耗时但操作简单，根据DNA片段大小，回收率在30%-90%之间。它可分别用于从中性和变性聚丙烯酰胺凝胶中分离双链和单链DNA。该方法广泛用于从变性聚丙烯酰胺凝胶中分离合成寡核苷酸。通过挤压浸泡从聚丙烯酰胺凝胶中回收的DNA通常适合用作杂交探针、聚合酶链反应（PCR）引物以及酶催化反应的底物。
