Here are 12 quick tips for NGS library preparation using NEBNext products.
Don't forget to spin down the vials before opening. This ensures everything is at the bottom of the vial and not in the cap or on the sides.
Make sure you use filter tips to minimize contamination.
Aseptic technique should always be used when working with RNA. Hands and dust particles may carry bacteria and mold and are the most common sources of RNase contamination. Make sure you clean your working surfaces with RNaseZap or a similar product and make sure you change your gloves frequently.
To further minimize contamination. Keep all sample and reagent vials capped when not in use.
Use RNase and DNase free plastics, water and solutions.
Keep isolated RNA on ice and avoid freeze thaw cycles. Purified RNA may be stored at minus 20 degrees or minus 70 degrees in water.
Be sure to use nuclease-free water for dilutions.
To efficiently mixed reaction components, gently pipette up and down and do not vortex.
Thaw and keep all master mixes on ice to ensure optimal activity.
When mixing reagents before addition to your reaction, keep everything on ice and use immediately after mixing.
Be sure to return enzymes to the freezer after use to ensure product stability.
To ensure the stability of your final library, store your samples in TE buffer and in low bind DNA tubes.
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