RNA sequencing has become a routine technique in modern genomics research. Yet for many laboratories, RNA library preparation remains one of the most error-prone steps in the workflow.
Unlike RT-PCR, traditional RNA library preparation requires multiple reagent components, repeated master mix preparation, and frequent pipetting. Every additional step increases the risk of pipetting mistakes, sample-to-sample variation, and failed library construction—especially when processing dozens or hundreds of samples.
The Challenge
Researchers commonly encounter:
- Too many reagent components
- Complex master mix preparation
- Pipetting errors during multi-step workflows
- Difficult training for new users
- Poor reproducibility across operators
For high-throughput projects, these challenges become even more significant.
A Premixed Workflow That Feels Like RT-PCR
The Hieff NGS™ EvoMax RNA Library Prep Kit (Strand-specific, Cat#12340) simplifies library preparation by combining key enzymes and reaction components into ready-to-use modules.
Instead of preparing multiple reaction mixtures, users simply add one premixed reagent per step.
|
Operation Step |
RNA Library Preparation |
RT-PCR |
|
RNA Extraction |
Yes |
Yes |
|
gDNA Removal |
No |
Extra 3 reagents (ddH₂O, gDNA digester buffer, gDNA digester), 1x PCR |
|
RNA Fragmentation |
Extra 1 reagent (Frag/Prime Buffer), 1x PCR |
No |
|
Reverse Transcription |
Extra 1 reagent (1st Reaction Module), 1x PCR |
Extra 4 reagents (ddH₂O, Reverse Transcriptase, RT Buffer, RT Primers), 1x PCR |
|
Second Strand Synthesis |
Extra 1 reagent (2nd Reaction Module), 1x PCR |
No |
|
Adapter Ligation |
Extra 1 reagent (Ligation Reaction Module), 1x PCR |
No |
|
PCR Amplification |
Extra 2 reagents (Amplification Enzyme, Adapter Primers), 1x PCR |
Extra 4 reagents (ddH₂O, Amplification Enzyme, Amplification Primers, Quantified Template), 1x PCR |
|
Purification |
Bead Purification |
Gel Extraction |
|
Sequencing |
Next-Generation Sequencing (NGS) |
Sanger Sequencing |
|
Downstream Steps |
None |
Vector Construction, etc. |
The result is a workflow that feels remarkably similar to RT-PCR:
Add one reagent → Run PCR → Continue to the next step.
Only five simple additions are required to generate a sequencing-ready RNA library.
Designed to Reduce Human Error
The premixed design helps laboratories minimize common experimental mistakes, including:
✓ Missing reagents
✓ Incorrect reagent combinations
✓ Inconsistent master mix preparation
Whether preparing a few samples or hundreds, the workflow remains consistent and easy to follow.
Proven Across Species and Platforms
1. RNA Library Preparation Across Different Species
Figure: Consistent library size distribution across multiple species. mRNA library preparation across different species, with the main peak of the libraries consistently centered around 500 bp.
2. Compatibility with Both Automated and Manual Library Preparation
|
RNA Standard |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
MGISP-960 Automated |
27.2 |
24.2 |
25.4 |
24.4 |
23 |
21.4 |
24.4 |
23.4 |
23.2 |
23 |
25.6 |
21.8 |
|
Manual |
26.2 |
24.6 |
26.2 |
25.3 |
22.8 |
20.5 |
25.1 |
22.5 |
23.6 |
23.9 |
24.6 |
20.9 |
|
Automated/Manual Yield (%) |
103.82% |
98.37% |
96.95% |
96.44% |
100.88% |
104.39% |
97.21% |
104% |
98.31% |
96.23% |
104.07% |
104.31% |
Figure: Comparison of automated and manual library preparation results on the MGI platform, showing consistent library profiles and comparable yields.
Related Products
|
Category |
Name |
Cat. No. |
Size |
|
|
RNA Lib Prep |
Dual-mode(Strand specific & Non Strand specific) |
12308ES24/96 |
24 T/96 T |
|
|
Premix version |
12340ES24/96 |
|||
|
12341ES24/96 |
||||
|
mRNA isolation |
Eukaryotic mRNA |
12629ES24/96 |
24 T/96 T |
|
|
rRNA depletion |
Human/Mouse/Rat |
Hieff NGS™ MaxUp Human/Mouse/Rat rRNA Depletion Kit(rRNA ITS/ETS) |
12257ES24/96 |
|
|
Hieff NGS™ MaxUp Human/Mouse/Rat rRNA Depletion Kit(rRNA ITS/ETS) 2.0 |
12726ES24/96 |
|||
|
Plant |
12254ES24/96 |
|||
|
Magnetic Beads |
- |
12602ES03/08/56 |
1/5/60 mL |
|
|
|
12601ES03/08/56 |
1/5/60 mL |
||