Promega
Wizard SV Gel and PCR Clean-Up System
A9282
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Using Wizard SV Gel and PCR Clean-UP sytem, the plasmid (8 Kb) restricted with restriction enzymes in a 50 microliter reactions were purified to remove enzymes and buffer components in the restriction reaction. Reactions were eluted in 30 microliter nuclease free water and recovery was 80% (820 ng from 1 microgram restriction) with very good quality (260:280 of 1.88). Purified plasmid backbone was further used for ligating with DNA insert.
Purification of DNA restriction reactions for cloning
1 microgram of plasmid DNA restricted with two restriction enzymes in a total volume of 50 microliter per prep
None
The protocol very easy to follow. Just follow the instructions comes with kit's instruction manual. I started with 1 microgram of my plasmid DNA restricted with two restriction enzymes (plasmid size was 8 Kb). Before proceeding further for ligation of DNA insert, I needed to remove restriction enzymes and other salts in the restriction reaction. Using the kit, I have got purified plasmid backbone with good quality of 260:280 ratio of 1.88. Recovery was 80 % (recoverd 820 ng from 1 microgram plasmid restriction reaction).
N/A
Less hands-on time perform the purification (less than 15 mintues)
Highly suggest this kit for purification of restriction reactions or PCR reactions for successful cloning method.