2×Hieff® Robust PCR Master Mix（With Dye)

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All-in-One Mix/2×Hieff® Robust PCR Master Mix（With Dye)

2×Hieff® Robust PCR Master Mix（With Dye)

This product is intended solely for scientific research use by professionals. It must not be used for clinical diagnosis or treatment, must not be used in food or pharmaceutical applications, and must not be stored in ordinary residential buildings.

Product Code	Product Specification	List Price	Promotional Price
12308	8T	445.00	445.00
12324	24T	1785.00	1785.00
12396	96T	3385.00	3385.00
12100	100T	15255.00	15255.00

Ribo-seq Kit from Geneseed is used for the preparation of high-throughput sequencing libraries, specifically designed for sequencing libraries of ribosome-protected mRNA fragments. This kit enables researchers to rapidly obtain high-quality Ribo-seq data through an efficient and simple workflow. The Geneseed Ribo-seq Kit is compatible with both MGI and Illumina high-throughput sequencing platforms, and supports both Full Adapter and Truncated Adapter types. The recommended RNA input range is 100 ng to 1 μg.

Ribo-seq technology is widely applied in research on cancer, metabolic diseases, and more, by sequencing ribosome-protected mRNA fragments to reveal details of gene expression regulation. Each reagent in the kit has undergone rigorous quality control and is suitable for experiments with low sample input, improving library preparation efficiency. Every batch of library preparation kits is validated through library construction and sequencing to ensure consistent performance and quality across batches.

The Geneseed Ribo-seq Kit must be used in conjunction with the rRNA depletion module and adapter kits.

High Sensitivity: Suitable for RNA samples with low input amounts, ensuring detection of low-abundance transcripts.
Fast and Convenient: Optimized workflow shortens library preparation time and improves experimental efficiency.
High Reproducibility: Ensures high consistency between samples, facilitating data comparison and analysis.
Broad Compatibility: Applicable to a variety of sample types, including animal cells, tissues, plants, fungi, and clinical samples.
Storage Condition: -20°C
Product Components:

Component No.	Component Name	8 Reactions	24 Reactions	96 Reactions	100 Reactions
12309	Nuclease	X	X	X	X
12398	mRNA Capture Beads	X	X	X	X
12387	Beads Binding Buffer	X	X	X	X
12376	Beads Wash Buffer	X	X	X	X
12365	Ligation Enhancer	X	X	X	X
12354	Novel T4 DNA Ligase	X	X	X	X

Usage Instructions:

1. This kit provides a complete ribosome profiling (Ribo-seq) solution, including reagents required for ribosome-protected fragment (RPFs) extraction, RNA fragmentation, library construction and purification, suitable for translatomics research.

2. Use fresh or liquid nitrogen flash-frozen cells (≥1×10^6 cells). After cell lysis, centrifuge to collect the supernatant. It is recommended to add cycloheximide (final concentration 100 μg/mL) to stabilize ribosomes.

3. Add 10 μL of RNase I (provided in the kit), incubate at 25°C for 45 minutes. Purify RPFs using the spin column provided in the kit, elute with 30 μL.

Frequently Asked Questions:

Q1: Possible reasons for low RPF yield?

Incomplete cell lysis: Optimize lysis buffer ratio or extend lysis time.

Low RNase I activity: Check enzyme storage conditions (avoid repeated freeze-thaw cycles) or extend digestion time.

Q2: How to resolve high rRNA contamination in sequencing data?

It is recommended to use an rRNA removal kit (e.g., Ribo-Zero) before library construction.

Check RNase I digestion efficiency to ensure ribosomes are not over-degraded.

Q3: Abnormal library fragment size (not 28 nt)?

Possible due to excessive RNA fragmentation: Shorten the reaction time at 70°C to 5–8 minutes.

Check T4 PNK enzyme activity to ensure complete end repair.

Q4: Which sequencing platforms are compatible with this kit?

Default adapter design is compatible with Illumina platforms (supports PE50/PE100). Adapters for other platforms can be customized upon request.

References:

Ingolia, N.T. et al. (2009). Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling. Science, 324(5924), 218–223.

Kit Optimization Guide: *[Attachment] RB-1000_Optimization_Guide.pdf*

Download the product instruction manual

Catalog Number	Product Name	Packaging
12309	Nuclease	XXXXXX
12398	mRNA Capture Beads	XXXXXX
12387	Beads Binding Buffer	XXXXXX
12376	Beads Wash Buffer	XXXXXX
12365	Ligation Enhancer	XXXXXX
12354	Novel T4 DNA Ligase	XXXXXX

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