Introduction

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The Purified DNA/RNA is used for RT-PCR and PCR detection.

Details

Specifications

Features	Specifications
Main FunctionsC	Co-isolation DNA and RNA from a single FFPE tissue sample
Applications	RT-PCR, cDNA synthesis, PCR and second-generation sequencing, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	No more than six 10µm sections of 150mm2 surface area or three 20µm sections of 150mm2 surface area.

Principle

FFPE samples are incubated in an optimized lysis buffer, which results in the release of RNA and precipitation of DNA. After centrifugation, the RNA-containing supernatant and DNA-containing pellet are then processed separately to purify RNA and DNA. For RNA purification, transfer RNA Lysate to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer. For DNA purification, transfer DNA Lysate to an adsorption column and DNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA was finally eluted with low-salt buffer.

Advantages

1.Use non-toxic dewaxing solution without contact with xylene
2.Obtain both DNA and RNA simultaneously from the same sample. Elute separately without affecting each other (Have the same steps and effects as top brand 80234, perfect substitute.)

Kit Contents

Contents	IVD5116
Purification Times	50 Preps
HiPure DNA Micro Column	50
HiPure RNA Mini Column I	50
2ml Collection Tubes	150
Proteinase K	50 mg
Protease Dissolve Buffer	5 ml
Buffer DPS	60 ml
Buffer FRL	15 ml
Buffer ATL	15 ml
Buffer RLC	15 ml
Buffer AL	15 ml
Buffer VHB	44 ml
Buffer RW2	25 ml
RNase Free Water	10 ml
Buffer AE	10 ml

Storage and Stability

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

Experiment Data

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The Purified DNA/RNA is used for RT-PCR and PCR detection.

The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.

NGS Single Stranded DNA Library Prep Kit workflow

Three index types are available for the kit:

Non-index (Cat.# 30081): Libraries do not have index.

Index(Cat.# 30082): Each of the index primers has a unique 6-base index sequence that can be used to identify libraries. ssDNA library multiplexing is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30083): ssDNA sample multiplexing up to 96 libraries is possible with unique dual indexes. We have developed a Four-Base Difference Index System. This makes it possible to generate indexes with at least 4 bases different from each other in the 8 bases index length. The unique dual indexing primers remove NGS errors (example: de-multiplexing errors, read mis-assignment, index hopping etc). Index information can be downloaded here.

Kit advantages:

• • • Quick and simple Protocol
      • Quick: Total protocol time is only 1.5 hours
      • Simple: Hands-on time only 10 minutes
    • Easy procedure based on:
      • The ready-to-use master mix: Made reaction setup easy
      • Less reaction components: Simplify the reaction preparation
    • Less magnetic beads needed: Reduced more than 50% of the beads for cleanup steps
    • Directional library
    • Single stranded DNA as input: From 50 ng to 500 ng

NGS Single Stranded DNA Library Prep Kit has similar library conversion efficiency and yield as compared to a regular DNA library prep kit.

Alignment rate and duplication rate: comparison of single stranded DNA library kit versus double stranded DNA library prep kit.

The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.