Qiagen
QIAquick Gel Extraction Kit (50)
28704
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I used Qigen's QIAquick Gel extraction kit to extract restriction enzyme digested pBAD plasmid backbone for cloning from 1% agarose gel. After extraction, recovery was 75 to 80% among three different reactions of RE digestion. This was sufficient amount of recovery routine cloning experiments. Quality and Quantity were great after checking with Nanodrop ND1000 spectrophotometer (yield was 120 ng/microliter and 260:280 ratio were 1.81 to 1.85).
Extraction of RE digested plasmid backbones from Agarose gel for cloning in S.enterica
NcoI and BamHI digested pBAD plasmid vector
140 ng/microliter of pBAD vector in a 20 microliter reaction were digested with RE enzymes and ran in the 1% agarose gel. Then excised intended back bone of pBAD from the gel and weighed into sterile eppendorf tube. Then performed gel extraction procedure as per kit's instruction. Used 4 times the volume of binding buffer to extracted band weighed in to eppendorf tube. Finally eluted using nuclease free molecular biology grade (pH 7.5) water rather than elution buffer which came with kit.
I prefer to use nuclease free molecular biology grade water to elute plasmid DNA at the end of the protocol.
Good quality recovery of pBAD plasmid vector after following QIAquick Gel extraction kit. I got around 75 to 80% recovery which were sufficient for my cloning experiments.
None
Simple protocol to follow. Less time on hands to perform.
I like Qiagen kits for DNA work and recommend this kit for gel extraction.
Product Name: QIAquick Gel Extraction Kit (50)Supplier Page