Manual

Product Number:PCK85M

Shipping and Storage

-20°C,dry ice transport.

Components

Component	PCK85M  (24rxns)	PCK85MM(96rxns)
ERAT Mix	48μL	192μL
10×ERAT Buffer	120μL	480μL
T4 DNA Ligase	72μL	288μL
T4 DNA Ligase Buffer	336μL	672μL×2
2×PCR Mix	1.2mL	1.2mL×2
DNA Control(50ng/μL,200bp)	20μL	20μL

Features

1. End repair and phosphorylation and adding A to complete in one step;
2. After the end repair without purification,directly add the Adaptor;
3. Ultra fidelity amplification,the maximum extent to reduce the amplification bias;
4. The library is suitable for multiple sequencing platforms:MGI sequencing platform:MGISEQ-2000, MGISEQ-200, BGISEQ-500 and other MGI sequencing platform lllumina GAllx, HiSacnSQ, HiSeq 250/2000/1000, MiSeqsequencing and other lllumina platform sequencers;
5. It is suitable for preparing gDNA and cDNA libraries after physical interruption.

Application

This kit is suitable for both Illumina and MGI sequencing platforms. It provides the premixed enzyme modules required for DNA end repair,5'-end phosphorylation modification,3'-end A addition and Adaptor ligation in DNA library preparation. It can be used with different sequencing platform adapters. Primer kits can prepare DNA into specific DNA libraries for Illumina or MGI sequencing platforms. Using high-fidelity DNA polymerase for library enrichment and unbiased PCR amplification,the coverage area of the sequence is expanded,and high-quality DNA libraries can be prepared. All reagents provided in the kit have undergone strict quality control and functional verification to ensure the stability of library preparation to the greatest extent.

Self-provided reagents and consumables

1. Magnetic rack:DynaMagTM-2 is recommended;
2. Anhydrous ethanol (100% ethanol,analytically pure);Deionized water (pH 7.0-8.0);
3. Adaptor index kit:

MGI platforms.

llumina platform.

• DNA purification recycling kit;

Reaction tube:Low adsorption PCR tube and 1.5mL centrifuge tube are recommended;

Tips:It is recommended to use high quality filter gun head to prevent contamination of kit and library samples.

Preparation and important things before the experiment

1. In order to avoid repeated freezing and thawing of reagents affecting the yield of the library,it is recommended to store the reagents in separate packaging when they are used for the first time;
2. Due to improper operation of PCR products,it is easy to pollute,resulting in inaccurate experiment results.It is suggested to isolate the preparation area of PCR reaction system from the purification area of PCR products,and use a special pipette to clean each experiment area regularly;
3. Sample preparation
   1. The fragment size of DNA sample should be concentrated:magnetic bead double selection can be carried out when the fragment of interrupted product is more dispersed;
   1. The recommended number of library cycles for this kit is adjusted according to the amount of input. For specific schemes,please refer to the instructions of each platform;
   1. The input of cfDNA must be greater than 1ng.
4. Reagent preparation
   1. Take out the corresponding reagent in the kit,centrifuge briefly,and put the enzyme mixture on ice for use:before use,dissolve the buffer solution at room temperature and then centrifuge it in shock,put it on ice for use,and put deionized water at room temperature for use; Please make the mixture on the ice.
   1. The buffer solution in the kit may precipitate after freezing and dissolving. The precipitation will not affect the function of the reagent. Please fully shake and mix until the precipitation disappears.

Self-provided reagents and consumables