Introduction

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from bacteria, yeast cells
Applications	RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Bacteria, yeast cells
Sample amount	Bacteria: <10^9；Yeast cells：<2 x10^7
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

Advantages

• Fast – several samples can be extracted in 30 minutes
• High purity – the purified RNA can be directly used in various downstream applications
• High recovery – RNA can be recovered at the level of PG
• Good repeatability – silica gel column purification technology can obtain ideal results every time
• Broad spectrum – it can deal with various bacteria, including Gram-positive bacteria that are difficult to be lysed
• Sufficient components – the kit contains carried wall breaking enzymes and glass beads

Kit Contents

Contents	R418202	R418203
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
Glass Beads (0.1-0.6mm)	30 g	150 g
DNase I	600 μl	5 x 600 μl
DNase Buffer	6 ml	30 ml
Protease Dissolve Buffer	1.8 ml	15 ml
Buffer ATL	50 ml	200 ml
Buffer PHC	50 ml	200 ml
Buffer GXP2*	20 ml	100 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Buffer PHC should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Buffer PHC up to 12 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.

The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic approach (for example, BioDynami DNA fragmentation enzymes, Cat.# 40061 and Cat.# 40062) and mechanical approaches such as sonication and nebulization etc.

NGS Low Input DNA Library Prep Kit Workflow

Three index types are available for the NGS Low Input DNA Library Prep Kit of illumina platform:

Non-index (Cat.# 30022): Libraries do not have index.

Index (Cat.# 30024): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify the low input DNA libraries. Library multiplexing up to 48 samples is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30025): Library multiplexing up to 96 low input DNA libraries is possible with unique dual indexes with our Four-Base Difference Index System. The system allows us to make indexes for the libraries that have at least 4 bases different from each other in the 8-base barcode length. Our unique dual index primers can reduce sequencing errors such as de-multiplexing errors, amplification errors, mis-assignment of reads, index cross-contamination, and also index hopping. The kit  includes 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34024).

Kit advantages:

• • • Fast protocol
      • The hands-on time is only 10 minutes
      • The total protocol time is around 1.5 hours
    • Simple procedure
      • Ready-to-use master mix makes it simple for reaction setup
      • Less reaction components
    • Less magnetic beads required for cleanup steps: Save the cost more than 50%
    • Low input DNA amount: Starts from 1 ng of DNA

Comparison of library conversion efficiency under the same NGS library preparation condition. Input DNA amounts are 1 ng and 10 ng, respectively. DNA was mechanical sheared with Covaris before library prep. BioDynami kit: NGS Low Input DNA Library Prep Kit (Cat. #30022).

Comparison of library yield under the same NGS library prep condition. Input DNA amounts are 1 ng, 10 ng, and 100 ng. DNA was mechanical sheared with Covaris before library prep. BioDynami kit (Cat. #30022). PCR cycle numbers were indicated.

• Set of three Solid Phase Adsorption Toxin Tracking (SPATT) Bags
• Pre activated
• Ready for deployment
• HP20

Description

Solid Phase Adsorption Toxin Tracking (SPATT) is a biomimetic in-situ water monitoring tool that falls under an expanding umbrella of passive samplers. It serves to warn researchers of toxin-producing harmful algal bloom (HAB) developments early on. It has been popularized through its affordability, ease of use, and its ability to capture ephemeral events in marine, brackish, and freshwater environments. Its uptake of contaminants has been shown to be more similar than other sampling methods to that of aquatic species like bivalves, mussels, and clams. It provides an average bioavailable fraction of a toxin over deployment time that can be used to determine an overall toxin risk to organisms. The sampling period typically depends on the bioactivity at a site, ranging from 24 hours to 4 weeks in most cases.

A SPATT passively absorbs and desorbs extracellular compounds over its stretch of time at a sampling site; in an organism, a toxin would go through biochemical detoxification processes. Passive samplers have a higher sensitivity for more compounds and provide improved stability and preservation of these compounds within the resin. SPATT devices capture less commonly detected cyanotoxins (e.g. cylindrospermopsin) at lower concentrations than that of a grab sample (collected at one point in time). Grab samples are limited in scope and sensitivity, and underrepresent toxins like microcystin-LR, which is picked up very reliably through SPATT technology.

Uses HP20 that is widely applicable for many toxins.

Used to capture:

• Cyanotoxin (e.g. microcystin and cylindrospermopsin)
• Saxitoxin & derivatives (GNTXs, C-toxins), and other paralytic shellfish toxins (PSTs)
• Domoic acid (DA)
• Nodularin
• Anatoxin-a
• Brevetoxins (via “red tide”-causing dinoflagellate Karenia brevis)
• Okadaic acid (OA) & derivative Dinophysistoxins (DTXs) and other Diarrheic shellfish poisoning (DSP) toxins
• Yessotoxin (YTX) and Pectenotoxins (PTXs)
• Cyclic imines (CIs), e.g. Spirolides (SPXs), Gymnodimines (GYMs), Pinnatoxins (PnTXs)
• Ciguatoxin (CTX) & precursor Gambiertoxin (GTX), Maitotoxin (MTx), and other ciguatera fish poisoning (CFP) toxins

Set of three Solid Phase Adsorption Toxin Tracking (SPATT) Bags
Pre activated
Ready for deployment
HP20