StarLighter Script III First-Strand Synthesis Kit

• The yield of reverse transcription products is higher;
• Obtain long cDNA fragments faster;
• Wider range of template applications.

• Single-strand cDNA synthesis;
• The synthesized single-strand cDNA can be widely used in 2ndStrand cDNA synthesis (library construction), hybridization, PCR/qPCR amplification, etc.

Product components


Product Performance

• Transcription results accurately reflect template quantity information

Reverse transcription was performed with a starting amount ranging from 0.2 ng to 2000 ng, and quantification was performed by qPCR. There was a good linear relationship between the log value corresponding to the reverse transcription template amount and the qPCR Ct value.

Fig.1 Comparison of qPCR results of reverse transcription products using different reagents

Reverse transcription was performed using 0.2 ng-2000 ng of human Hela cell line RNA (DNA removed) as a starting template. 1/10 volume of the reverse transcription product was used as a quantitative template, and quantification experiments were performed using Starlighter SYBR Green qPCR Mix (YWHAZ primers, product 249 bp, product GC content 44.6%). The log values corresponding to the reverse transcription template were plotted against the qPCR Ct values. Starlighter, TA2, and ABIII showed very good linear relationships, and the amplification efficiencies were all between 95-110%.

• High reverse transcription efficiency. The GC content is 60-75%. Qihengxing reverse transcription reagent and AB-III show good reverse transcription efficiency in the range of 4 ng-2000 ng.

Fig. 2 Comparison of reverse transcription efficiency of different reagents. The reverse transcription efficiency of 1000 ng RNA input was set as 100%. The relative reverse transcription efficiency of each kit at different input amounts was calculated based on the experimental results.

• Easily handle reverse transcription and quantification of GC-rich templates

For reverse transcription and quantification of a gene with a GC content of 75.5% (human ApoE), the Qihengxing reverse transcription kit achieved an excellent linear relationship between input and output, R ² =1.

Fig.3 Comparison of quantification results of high GC fragments by qPCR quantification of products transcribed using different reagents

Reverse transcription was performed using varying amounts of DNA-depleted human Hela cell line RNA as a template. 1/10 volume of the reverse transcription product was used as a quantitative template for quantification using the StarLighter SYBR Green qPCR Mix (ApoE primers, product 322 bp, GC content 75.5%). The linear relationship between the amount of reverse transcribed RNA and the qPCR Ct value was compared using different reverse transcription reagents.

• Easily handles degraded and low-input samples

Using RNA with different degrees of degradation as templates - continuous gradient dilution (20-0.2ng), it can be effectively amplified.

• Compatible with various types of samples

Compared with suppliers TA2 and TS, the Qihengxing reverse transcription kit showed good performance in a variety of different samples and had lower Ct values.

Fig. 4. Reverse transcription of RNA from different species using different reagents and quantification of the products. Reverse transcription was performed using 200 ng of RNA from different species as template. A 1/10 volume of the reverse transcription product was used as the template for quantification. Quantification was performed using the StraLighter SYBR Green qPCR Mix, and the Ct values of the different reverse transcription reagents were compared.