StarLighter Script IV First Strand Synthesis Kit

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

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

• Transcription results accurately reflect template quantity information

Reverse transcription was performed with a starting amount ranging from 0.2ng to 2000ng, and quantification was performed using 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 amount as a template. 1/10 volume of the reverse transcription product was used as a quantitative template, and the quantitative experiment was performed using Starlighter SYBR Green qPCR Mix (YWHAZ primers, product 249 bp, product GC content 44.6%). The log value corresponding to the reverse transcription template was plotted against the qPCR Ct value. Starlighter, TA2 and ABIII showed a very good linear relationship, 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%, and the relative reverse transcription efficiency of each kit at different starting amounts was calculated based on the test results.

• Easily handle reverse transcription and quantification of high GC 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. ² =1.

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

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

• Easily handles degraded and low-input samples

Using RNA with different degrees of degradation as templates - continuous gradient dilution (20-0.2ng), effective amplification can be achieved.

• 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 Different reagents were used to reverse transcribe RNA from different species and the products were quantified. 200 ng of RNA from different species was used as template for reverse transcription, 1/10 volume of reverse transcription product was used as quantitative template, and the quantitative experiment was performed using StraLighter SYBR Green qPCR Mix to compare the Ct values of different reverse transcription reagents.