Graph 1: Eluting 12 kb pCAMBIA plasmid with preheated buffer at 70 ˚C significantly improves DNA recovery compared to room temperature elution. Preheated buffer enhances total DNA yield by up to 25%.

Graph 2: Increasing the RNase A concentration from 0.13 mg/mL (PrimeWay I) to 0.17 mg/mL (PrimeWay II) improved both DNA yield and purity. The A₂₆₀/₂₈₀ ratio improved from 2.11 to 1.88, demonstrating elimination of RNA contamination and purer plasmid DNA.

Figure 1: RNase A is provided with higher volume and packed in a 1.5 mL microcentrifuge tube.

Table 2: Comparison of RNase A volumes between PrimeWay I and PrimeWay II

Figure 3: The data shown from 3 graphs above indicate the PrimeWay Plasmid II Kit efficiently extracts high-quality plasmid DNA across different sample types. The consistent A260/280 and A260/230 values indicate that the protocol minimizes contamination and preserves DNA integrity, making it suitable for sensitive downstream applications such as restriction digestion, cloning, or qPCR.

Graph 3: 5 mL overnight grown culture of various plasmid sizes with different high copy number are extracted using different brands. Plasmid extraction is performed according to manufacturing protocol and eluted with 100 µL elution buffer.

Graph 4: 5 mL overnight grown culture of various plasmid sizes with different low and medium copy number are extracted using different brands. Plasmid extraction is performed according to manufacturing protocol and eluted with 100 µL elution buffer.

Figure 4: A comparison of 100 ng plasmid DNA extracted using PrimeWay II and competitor brands P and Q was performed on a 1% TAE agarose gel, stained with Floro+Red Nucleic Acid Stain (BIO-5171-600 µL). The ExactMark 1 kb DNA Ladder (1st BASE, BIO-5140-50 µg) was used as the molecular size reference. The results demonstrate that PrimeWay II delivers plasmid DNA of comparable quality to the competitor brands.

Figure 5: The pCAMBIA plasmid (12 kb), extracted with the PrimeWay II Plasmid Kit, was successfully digested using common restriction enzymes. Clear bands and expected fragment patterns confirm the DNA high purity and suitability for subcloning and mapping.

Figure 6: The pJET-BRCA plasmid, extracted using the PrimeWay II Plasmid DNA Extraction Kit from E. coli 10β cells, was prepared for direct use in Sanger sequencing. Sequencing was performed using BigDye™ Terminator v3.1 chemistry, yielding a typical long, high-quality read consistent with automated capillary sequencing standards. This demonstrates the effectiveness of PrimeWay II in producing plasmid DNA of sufficient purity and integrity for precise downstream applications.