How to resuspend eBlocks and gBlocks Gene Fragments
Standard gBlocks Gene Fragments and gBlocks HiFi Gene Fragments are dried down and shipped in tubes (unless otherwise requested to ship in plates or ship wet). Plate products, including gBlocks and eBlocks Gene Fragments, are shipped on dry ice, if wet and ambient, if dry. Once you receive your DNA fragments, it’s recommended to perform IDT’s resuspension protocol. gBlocks Gene Fragments are stable for up to two years while dried and stored at −20C.
Follow the steps below when resuspending your DNA fragments:
1. Prepare your DNA fragment: Once you receive your DNA fragments, spin down the tube in a microcentrifuge for 5 seconds. This ensures that the DNA pellet is at the bottom. Since the pellet can be statically charged or lodged in the cap during shipping, doing this step prevents the pellet from flying out of the tube or remaining in the cap thus losing yield.
2. Add molecular grade water or buffer: After centrifugation, resuspend the pellet in molecular-grade water or TE buffer (pH 7.5–8) the required concentration. We recommend you use either a molecular grade water or a buffer such as IDTE, pH 8, to achieve a final concentration of 10 ng/µL. Please note that concentrations <1 ng/µL can result in material loss due to adherence to the plastic tube.
3. Vortex briefly: Mix the solution briefly using a vortex.
4. Heat the tube: Incubate at approximately 50°C for 15–20 minutes. Heating ensures the solvent contacts the entire pellet, even if it is stuck to the side of the tube, thereby increasing the likelihood of complete resuspension.
5. Vortex and centrifuge: After heating, briefly vortex again and then centrifuge.
6. Measure final concentration: Verify that the concentration of the resuspended gBlocks Gene Fragments to ensure that the full product has gone into solution. Use Nanodrop quantification to verify the final concentration. This method is preferred for short gBlocks Gene Fragments, as the Qubit™ system may give artificially low readings[1].
How to store eBlocks and gBlocks Gene Fragments
IDT Gene Fragments are stable for up to two years under proper storage conditions.
Here are some recommendations for improving the preservation of your gBlocks Gene Fragments:
- After resuspending in a high-quality, molecular-grade water or buffer, pH 7.5–8.0, you should store the DNA at −20°C and if necessary, aliquot to avoid more than 2 or 3 freeze-thaw cycles. Adding a carrier to your resuspension and dilution buffers can help prevent any potential decrease in product concentration.
- If stored dry at ambient temperature, IDT Gene Fragments remain stable for approximately three months due to inherent moisture causing DNA damage over time.
- It’s recommended that you generate aliquots at concentrations >1 ng/µL and reduce repetitive freeze-thaw cycles of the stock. Avoid more than 3 freeze-thaw cycles. If necessary, aliquot your IDT Gene Fragments before freezing to minimize degradation.
How to stabilize eBlocks and gBlocks Gene Fragments
If storing the DNA fragments at a concentration less than 1 ng/µL, you might experience a decrease in your final concentration over time. We have observed a decrease in DNA fragment concentrations in solutions with a starting concentration of less than 1 ng/µL, even when stored in low-bind tubes. This may be related to the very high purity of DNA fragments. Synthetic DNA lacks normal cellular debris found in DNA isolated from natural sources. In the absence of these natural carriers, synthetic DNA will bind irreversibly to plastic, resulting in DNA loss. Hence, small amounts of carrier can prevent this loss.
To prevent concentration loss, consider adding natural carriers like tRNA or poly A (0.1–1.0 mg/mL) to prevent product loss over time. You can add tRNA or poly A at concentrations of 0.1–1.0 mg/mL to the dilution buffers. Both tRNA (55714-250MG) and Poly A (P9403-25MG) can be sourced from Sigma. Without these natural carriers, the gBlocks Gene Fragments bind irreversibly to the tube plastic over time, leading to the observed decrease in concentration. Adding a carrier to your resuspension and dilution buffers can help prevent this loss of product.
By following these guidelines, you can ensure the integrity and longevity of your gBlocks Gene Fragments. Ready to use your gBlocks Gene Fragments for your experiment? Read more about how to clone with your IDT Gene Fragments by downloading our free cloning guide.
