Preparation of libraries for Next Generation Sequencing can be challenging for newcomers, often involving multi-step protocols which may stretch over several days. When planning the different steps of library preparation, basic bench rules and molecular biology techniques often get "lost" among all the other considerations.

Below, we have compiled some tips & tricks to make sure you get these basics right.

  1. Don't forget to spin down the vial before opening. This ensures your precious reagents are at the bottom of the vial and not in the cap or on the sides of the vial.
  2. Make sure you use filter tips to minimize contamination and change the tip after every pipetting step.
  3. Make sure you clean your working surface with DNA removal agent, such as DNA-ExitusPlus (OJ14818.10) and make sure you change your gloves frequently.
  4. Use good pipetting technique. To efficiently mix reaction components, gently pipette up and down and do not vortex.
  5. Always make a mastermix. Thaw and keep all master mixes on ice to ensure optimal activity.
  6. When mixing reagents before addition to your reaction, keep everything on ice immediately after mixing.
  7. Take care with your bead-based cleanups. Always use freshly prepared ethanol solutions for bead washes.
  8. Be sure to return enzymes to the freezer after use to ensure product stability.
  9. To ensure the stability of your final library, store your final libraries in TE buffer and in low-bind DNA tubes.
  10. Keep good records. Write down lot numbers of enzymes or kits and keep your notes if you're following a kit protocol.
Tech tips

LubioScience's NGS library prep kits

Using high-quality NGS library prep kits is key to producing high-quality sequencing data. 

With xGen NGS, we’ve made library preparation easy. IDT's solutions were developed to improve your time, yields, and budget across a broad range of sample types. Wherever your research journey takes you, you can trust in xGen NGS to assist.

Please do not hesitate to contact us for further assistance in choosing the suitable solution for your application needs.

More about DNA library prep kits More about RNA library prep kitsMore about adapters for NGS

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Integrated DNA Technologies (IDT)

Integrated DNA Technologies (IDT) is your advocate for the genomics age. They produce tools for NGS, CRISPR, qPCR and PCR. Their products include DNA/RNA oligos, genes and gene fragments. For more than 30 years, IDT's innovative tools and solutions for genomics applications have been driving advances that inspire scientists to dream big and achieve their next breakthroughs. 

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