GENVEYOR

Lipofectamine

Flow cytometry scatter plots for Lipofectamine-delivered GFP circular ssDNA at 100 and 250 ng, with GFP-positive populations of 0.42% and 0.24% — almost no cells above the gate.
Jurkat T cells, GFP-encoding circular ssDNA delivered with Lipofectamine. GFP-positive 0.42% (100 ng) and 0.24% (250 ng). Axes SSC-A versus BL1-A::GFP-GFP-A.

AccuLNP™

Flow cytometry scatter plots for AccuLNP-delivered GFP circular ssDNA at 100 and 250 ng, with GFP-positive populations of 23.4% and 33.2% — a large shifted population above the gate.
Jurkat T cells, GFP-encoding circular ssDNA delivered with AccuLNP™. GFP-positive 23.4% (100 ng) and 33.2% (250 ng) — 56× and 138× the matched Lipofectamine control on the left. Axes SSC-A versus BL1-A::GFP-GFP-A.

AccuLNP™ p-DNA / Circular ssDNA Transfection Kit

Catalog#
NTP-0200
Size
160 reactions for 24-well plates
Price (USD)
$229

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  • pDNA
  • Circular ssDNA

Product Overview

AccuLNP-DNA is a cell transfection reagent mediated by lipid nanoparticles (LNP) for DNA transfection. AccuLNP can efficiently interact with DNA molecules to form (lipid molecules + DNA) complexes and can effectively release DNA molecules within cells to achieve their biological functions. This reagent has the characteristics of high transfection efficiency, low cytotoxicity, and easy operation.

Validated cell lines HEK293 · CHO-k1

Product Detail

Catalog# Name Qty
NTP-0200-L Reagent L (liquid) 1 (250 µL)
NTP-0200-A Reagent A (liquid) 1 (250 µL)
NTP-0200-ST Reagent ST (liquid) 1 (10 mL)
Storage condition
-20 °C
Shelf life
12 months
Encapsulation time
~60 s
Cell viability
>90%
Format
Three-reagent liquid kit — Reagent A, Reagent L, Reagent ST

Protocol

Table 1. Recommended conditions for cellular transfection with the AccuLNP-DNA kit.
For one well 48-well plate 24-well plate 12-well plate 6-well plate
Surface area 1 cm²1.9 cm²3.8 cm²9.6 cm²
Complete growth medium (when plating cells) 250 µL500 µL1 mL2.5 mL
DNA (0.2 µg/µL stock) 0.25 µg — 1.25 µL0.5 µg — 2.5 µL1 µg — 5 µL2.5 µg — 12.5 µL
Reagent A 0.625 µL1.25 µL2.5 µL6.25 µL
Reagent L 0.625 µL1.25 µL2.5 µL6.25 µL
Reagent ST (to dilute transfection complex) 25 µL50 µL100 µL250 µL

The following procedure describes how to perform transfections in a 24-well plate. The surface areas of other cell culture vessels are different, and transfection should be scaled accordingly. Follow Table 1 to appropriately increase or decrease the amount of reagent use.

  1. Plate cells

    1. Approximately 18–24 hours before transfection, plate cells in 0.5 mL complete growth cell culture medium per well in a 24-well culture plate. Ideally cells should be at least 70% confluent prior to transfection. For adherent cells, plate at a density of 0.8–3.0 × 10⁵ cells/mL; for suspension cells, 2.5–5.0 × 10⁵ cells/mL.

    2. Incubate the cell culture overnight.

  2. Prepare the transfection complex

    1. Take the transfection kit out of the −20 °C freezer.

    2. Thaw Reagent A and Reagent ST in a 37 °C water bath.

      Note: Ensure Reagent A and Reagent ST are completely thawed and clear transparent solutions are obtained. Apply vortex mixing to the microfuge tubes of Reagent A and Reagent ST to ensure the solutions are homogeneous. Cool Reagent A and Reagent ST down to room temperature before use.

    3. Thaw Reagent L in a 37 °C water bath for at least 10 minutes. Apply vortex mixing to the microfuge tube of Reagent L to ensure a homogeneous solution is obtained. Cool Reagent L down to room temperature before use.

    4. Add 0.5 µg (2.5 µL of 0.2 µg/µL) DNA stock solution in a sterile tube.

    5. Add 1.25 µL Reagent A. Pipet gently to mix completely.

    6. Add 1.25 µL Reagent L to the above diluted DNA solution. Use a vortex mixer to mix quickly for at least 15 seconds. The resulting total volume is 5 µL.

    7. Add 50 µL Reagent ST to dilute the above transfection complex. Pipet gently to mix completely.

  3. Cell transfection

    1. Add the transfection complex prepared in Step 2 (total 55 µL) dropwise to different areas of the well.

    2. Gently shake the culture vessel to evenly distribute the transfection complexes.

    3. Continue culturing in the cell culture incubator for the required time (such as 24 hours), and then the user determines the detection method to evaluate the transfection efficiency.

Notes

  1. Unused solutions of Reagent A, Reagent ST and Reagent L can be stored at −20 °C. Thaw and dissolve again in a 37 °C water bath before next use.
  2. The recommended DNA stock solution to use is between 0.15 and 0.25 µg/µL. If the DNA stock solution concentration is higher than 0.25 µg/µL, dilute it with RNase-free water or appropriate buffer first to make sure the DNA concentration is within the above range, and then proceed to prepare the transfection complex.
  3. To determine the optimal reagent levels to use for DNA transfection, users can test a range of Reagent A levels (e.g. 0.625, 1.25, 1.875 µL for a 24-well plate) with a range of Reagent L levels (e.g. 0.625, 1.25, 1.875 µL for a 24-well plate). Following this approach, a total of nine different transfection complexes will be generated and transfected, with the aim to identify the best transfection condition for the specific DNA and cell type.

Ordering

Catalog number NTP-0200. Net 30 on an institutional purchase order. Prices exclude shipping and applicable tax. For scale-up, custom formats, or bulk pricing, email contact@genveyor.com.