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Plant Molecular Biology

High Throughput RTPCR Kit


A Novel High Throughput Solution for Gel-Based Gene Expression profiling. The JumpStart RED HT RT-PCR Kit is designed for high throughput gel-based RT-PCR applications, optimized for end-point detection of target RNA in a complex sample matrix. The kit contains a combination of Sigma's enhanced avian myeloblastosis virus reverse transcriptase (eAMV-RT™) and JumpStart RedTaq™ DNA polymerase.
  • Greater transcription efficiency
  • Enhanced sensitivity and specificity
  • Direct sample loading after PCR
  • Maximum flexibility and minimum optimization
 
 
Product Code Description Quantity
J3520 JumpStart RED HT RT-PCR Kit 40 reactions
200 reactions
 
eAMV has superior performance and thermal stability
Sensitivity for low abundance message is critical for comparative gene expression studies. eAMV has superior processivity and thermal stability compared to other reverse transcriptases. These unique features make it the ideal enzyme for detection of low abundance RNA, especially those with difficult secondary structure.
Figure 1. RT-PCR was performed on total RNA-HELA cells serially diluted from 1000 ng to 1 ng. Reactions were performed with b actin primers according to suppliers' protocols.
Lanes 1, 10, 11, 20: PCR Marker
Lanes 2, 6, 12, 16: 1000 ng
Lanes 3, 7, 13, 17: 100 ng
Lanes 4, 8, 14, 18: 10 ng
Lanes 5, 9, 15, 19: 1 ng
 
Enhanced sensitivity and specificity with JumpStart Taq DNA Polymerase
The addition of JumpStart antibody to the PCR mix increases sensitivity and specificity of PCR amplification in both the one-step or two-step reactions.
Figure 2. RT-PCR amplification of a 440 bp fragment from tobacco mosaic virus RNA template was performed using the one-step protocol.
Lane 1: DNA ladder
Lanes 2 & 3: 2 ng starting RNA
Lanes 4 & 5: 200 pg starting RNA
Lanes 6-7: 20 pg starting RNA
 
Direct loading of PCR product
The specially formulated PCR mix containing a red tracking dye allows easy setup of PCR reactions and direct sample loading after PCR. This feature saves time and lab supplies, especially for high throughput gel-based RNA detection.
Figure 3. Left panel shows PCR setup: no enzyme added, addition of enzyme, and completely mixed RT-PCR reaction. Right panel shows direct loading of PCR products. expression profiling of multiple genes from a single RNA sample.
 
Maximum flexibility and minimum optimization
The JumpStart RED HT RT-PCR Kit includes all reagents for performing one-step and two-step RT-PCR. The one-step procedure is suitable for detection of a single gene from an RNA sample, whereas the two-step procedure is suitable for expression profiling of multiple genes from a single RNA sample.
Figure 4. Amplification of three Arabidopsis genes from five different tissues using one-step and two-step RT-PCR.