ZHIXUAN Reagent Series

ZHIXUAN  Universal DNA Library Prep Kit for MGI

Whole genome sequencing  |  Amplicon sequencing  |  Immunoprecipitation sequencing  |  Metagenomic sequencing  |  Methylation sequencing  |  16srDNA sequencing  |  Targeted sequencing

ZHIXUAN Universal DNA Library Prep Kit for MGI is a library prep kit specially optimized for the MGI high-throughput sequencing platform. This kit can convert 100 pg - 4 μg Input DNA into libraries dedicated for MGI high-throughput sequencing platform. As a new upgraded version,ZHIXUAN Universal DNA Library Prep Kit for MGI has significantly improved the library conversion rate and library amplification output through overall improvements to the end repair module, connection module and library amplification module. It is widely used in PCR library construction of various sample types. All reagents provided in the kit have gone through strict quality control and functional validation, which greatly guarantees the stability and repeatability of library construction

·  Shorter library prep time, one single library prep takes only 75 min

·  Higher adapter connection efficiency, higher library conversion rate

·  Easier and faster operation, premix reduces operation errors

·  Wider template compatibility range, initial Input DNA can be from 100 pg to 4 μg

·  Wider sample compatibility, suitable for fragmented DNA, cfDNA, FFPE DNA, ChIP DNA, etc.

The product shall be stored in environment of temperature range between -30°C to -15°C.

For transportation, temperature range between -20°C to 0°C is required.

FAQ

Q1: Low library output
A1: 
(1) Re-quantify the amount of input template to check whether the initial template amount is correct. If it is below the lower limit (100 pg), the template input amount shall be increased;
(2) If the template quality is poor, the number of cycles can be appropriately increased according to the IFU or high-quality templates can be used;
(3) Please check whether each step of the system and procedure is carried out in accordance with the IFU, and pay attention to the details of each operation in each step;
(4) The drying process of the library purification magnetic beads shall not be too long, which causes the magnetic beads to be too dry. Overdrying will also reduce the output;
(5) When conducting library amplification, ensure that there is no magnetic bead residues. Otherwise it may inhibit the amplification reaction;
(6) When quantifying the qPCR library, ensure that the cycle time and number of cycles are correct.

Q2: After library construction, the proportion of chimeras is high.
A2: This situation is due to over-amplification. Over-amplified libraries are non-specific amplification caused by direct mismatches between libraries when the primers have been exhausted, and unintended chimeras are introduced. 

Q3: The same index is added to different samples, how to solve it?
A3: Separate samples with the same index into different lanes for sequencing.

Q4: The Agilent 2100 detects a distinct peak characteristic of adapter dimers.
A4:
(1) The multiplier of magnetic bead purification can be appropriately reduced;
(2) The adapter concentration can be appropriately reduced.

Q5: Other operation notes
A5: 
(1) Before use, thaw each reagent of the kit at room temperature. After thawing, invert each reagent several times to mix thoroughly, centrifuge briefly, and place them on ice for later use;
(2) When preparing the reaction solution for each step, it is recommended to use pipettes to mix it properly. Intense shaking may cause decrease in library output.
(3) To avoid cross-contamination of samples, it is recommended to use pipette tips with filter. Please change the pipette tip when handling different samples.
(4) It is recommended to perform each step of reaction in a PCR instrument with a heating cover. The PCR instrument shall be preheated to near the reaction temperature before use.
(5) PCR products can easily lead to aerosol contamination due to improper operation, which will affect the accuracy of test results. Therefore, we recommend mandatory physical isolation between PCR reaction system preparation area and PCR product purification detection area, and use dedicated pipettes, regularly clean each experimental area (using 0.5% sodium hypochlorite or 10% bleach for wiping and cleaning) to ensure the cleanliness of the experimental environment.