YALEPIC^® Bacterial RNA Extraction Kit (with DNase I)

This kit is suitable for total RNA extraction from Gram-negative bacteria (e.g., E. coli, Salmonella, Pseudomonas aeruginosa) and Gram-positive bacteria (e.g., Staphylococcus aureus, Bacillus subtilis, Streptococcus). For Gram-positive bacteria with thicker cell walls, the lysis time can be appropriately extended or combined with lysozyme pre-treatment for optimal lysis efficiency and RNA yield.

This kit uses an optimized wash system that effectively removes the vast majority of genomic DNA during extraction. The kit also includes RNase-free DNase I, enabling on-column gDNA digestion. RNA products after digestion typically show no gDNA amplification bands by PCR testing, meeting the stringent gDNA carryover requirements of downstream experiments such as RT-qPCR and RNA-Seq.

No. The YALEPIC Bacterial RNA Extraction Kit uses a guanidine isothiocyanate lysis and silica-membrane spin column purification approach throughout, without requiring phenol, chloroform, or other toxic organic solvents for extraction. The operation is safer and more environmentally friendly, substantially reducing the risk of personnel exposure to toxic chemicals, and is particularly suitable for routine laboratory and high-throughput extraction settings.

Under skilled operation, total RNA extraction from bacterial samples using this kit takes approximately 30 minutes for the complete workflow (including lysis, binding, washing, and elution steps). If on-column DNase I digestion is performed, add approximately 15 minutes. When processing multiple samples in batch, parallel operations can further improve experimental efficiency.

Using E. coli as an example, 10-30 microgram of high-purity total RNA can typically be obtained from 1×10^8 cells. For RNA purity, the A260/A280 ratio is generally between 1.9 and 2.1, with A260/A230 ratio >=2.0, indicating extremely low protein and organic solvent carryover. Agarose gel electrophoresis clearly shows 23S and 16S rRNA bands, with a 23S/16S ratio >=1.8, demonstrating excellent RNA integrity.

The recommended bacterial input is 1×10^6 to 1×10^8 cells; the maximum is not recommended to exceed 1×10^9 cells. Too little sample may result in low RNA yield; too much sample leads to incomplete lysis and column overload, reducing RNA purity and recovery. It is recommended to perform a pilot experiment based on bacterial species and growth state to determine the optimal sample input.

The total bacterial RNA extracted with this kit is of excellent quality and has been validated for a wide range of downstream applications including: RT-PCR, qRT-PCR (gene expression analysis), Northern Blot/Dot Blot (RNA detection), transcriptome sequencing (RNA-Seq), gene chip analysis (Microarray), Poly(A) mRNA selection, in vitro translation, RNase protection assays, and molecular cloning (cDNA library construction).

DNase I should be stored sealed at -20 degrees C and avoid repeated freeze-thaw. It is recommended to aliquot DNase I into small volumes (e.g., one tube per use) upon receipt. Use one tube at a time and continue storing unused aliquots at -20 degrees C. Repeated freeze-thaw causes DNase I activity decline and affects gDNA digestion efficiency.

Common causes of RNA degradation and resolution suggestions: RNase contamination – ensure use of RNase-free tips, tubes, and work surfaces. Wear gloves and a mask throughout the operation. Delayed sample processing – bacteria should be lysed immediately after collection, or temporarily stored in RNA protection solution at -80 degrees C. Incomplete lysis – ensure lysis buffer is thoroughly mixed with bacterial pellet. For Gram-positive bacteria, extend lysis time or add a lysozyme treatment step. Elution buffer issues – be sure to use the kit-provided RNase-free Water for elution; avoid using non-RNase-free water.

Compared with similar products on the market, the core advantages of the YALEPIC Bacterial RNA Extraction Kit include: Safe and non-toxic – no phenol/chloroform extraction required throughout, safer operation. Included DNase I – contains DNase I and dedicated buffer, no additional purchase needed, all in one kit. High RNA quality – unique silica-membrane material ensures high RNA purity and integrity. Broad bacterial species compatibility – simultaneously suitable for both Gram-negative and Gram-positive bacteria. High cost-effectiveness – 50T specification at only 750 CNY, per-reaction cost of only 15 CNY; additional discounts for bulk purchase. Manufacturer direct supply – independently developed and produced by YALI BIOTECH, stable quality with timely technical support response.

This kit is designed for manual spin column operation. For high-throughput automated extraction, YALI BIOTECH offers a matching magnetic bead-based bacterial RNA extraction kit that is fully compatible with YALI BIOTECH’s automated nucleic acid extractors and other mainstream automated platforms, achieving high-throughput extraction of 96 samples per batch. Please consult our technical team for automated workflow recommendations.

YALI BIOTECH provides full professional technical support for all products. If you encounter any issues during use, you can obtain help through the following channels: Technical service hotline: 0512-66256010, Technical support email: info@yalibiotech.com, Official website online message: www.yalibiotech.com.