Introduction

The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 0.25ml body fluids using column and MagZol LS reagent
Applications	qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection
Purification method	Mini spin column
Purification technology	Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology)
Process method	Manual (centrifugation or vacuum)
Sample type	Blood, serum, plasma
Sample amount	250μl
Elution volume	≥20μl
Liquid carrying volume per column	800µl
Binding yield of column	100μg

Principle

0.25ml liquid samples are homogenized in 0.75ml MagZol LS Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100 µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30–100µl of RNase-free water.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – isolation of several samples can be completed in 40 minutes by using column purification method
• Safety – no phenol chloroform extraction required
• Sensitive – direct lysis of blood, plasma and other samples without separation of leukocytes

Kit Contents

Contents	R416302	R416303
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	2 x 125
MagZol LS Reagent	60 ml	270 ml
Buffer RWC	20 ml	60 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

MagZol LS Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.

Description 

The PM2800 ExcelBand™ 3-color Extra Range Protein Marker is a ready-to-use three-color protein standard with 13 pre-stained proteins covering an extra range of molecular weights from 10 to 310 kDa in Tris-Glycine buffer (9 to 290 kDa in Bis-Tris (MOPS) buffer and 10 to 290 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore except for three reference bands (one green and two red bands at 25 kDa and 75, 310 kDa respectively) when separated on SDS-PAGE in Tris-Glycine buffer. The PM2800 ExcelBand™ 3-color Extra Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (nitrocellulose, PVDF, or nylon) and for approximating the size of proteins. The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading.

Features

• Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
• Three reference bands — 310 kDa (red), 75 kDa (red) and 25 kDa (green)

Contents

Approximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25℃), 2 % SDS, 0.2 mM Dithiothreitol, 3.6 M Urea, and 15 % (v/v) Glycerol). 

Quality Control

Under suggested conditions, PM2800 3-color Extra Range Protein Marker resolves 13 major bands in SDS-PAGE (Tris-Glycine buffer, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. 

Storage

4°C for 3 months
-20°C for long term storage

The PM2800 ExcelBand™ 3-color Extra Range Protein Marker is a ready-to-use three-color protein standard with 13 pre-stained proteins covering an extra range of molecular weights from 10 to 310 kDa in Tris-Glycine buffer (9 to 290 kDa in Bis-Tris (MOPS) buffer and 10 to 290 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore except for three reference bands (one green and two red bands at 25 kDa and 75, 310 kDa respectively) when separated on SDS-PAGE in Tris-Glycine buffer. The PM2800 ExcelBand™ 3-color Extra Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (nitrocellulose, PVDF, or nylon) and for approximating the size of proteins. The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading.

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.

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.