Preparation of sample libraries for next-generation sequencing!
                     P R E PA R AT I O N  O F  S A M P L E  L I B R A R I E S  F O R  
N E X T- G E N E R AT I O N  S E Q U E N C I N G !
H T T P S : / / T E K M AT I C . C O M
Next-generation sequencing allows the profiling of 
sequences from transcriptomes and genomes and DNA-
protein interaction. The techniques used are an integral 
component of the research process and discoveries in the 
field of biology. The capability to quickly create large 
volumes of sequence information offers various 
applications, speeding advancements in research and 
transforming our understanding of the human body’s 
health and diseases.
P R E PA R I N G  S E Q U E N C I N G  
L I B RA R I E S  F O R  S E Q U E N C I N G
For DNA, or cDNA (synthesized using the RNA), to be sequenced, it 
must be separated, repaired at the end, and put into libraries for 
sequencing. Put the term sequencing libraries refers to pools of 
DNA fragments containing adapter sequences that work with a 
particular sequencing platform and indexing barcodes that allow for 
identifying unique samples. The primary methods for library 
preparation include ligation-based library prep, segmentation-based 
library preparation, and amplicon library preparation. The particular 
protocol you select depends on your sequencing platform and the 
downstream analysis. The most fundamental steps for library 
preparation include fragmentation, end repair, as well as the 
addition of adapters, and (optional) Amplification of PCR:
End repair and fragmentation Short-read sequencing 
techniques like Illumina cannot easily analyze very long DNA 
strands. As a result, DNA must be broken down into smaller, 
uniform pieces. After the fragmentation process, the DNA 
fragments are either repaired or polished at the end. A single 
Adenine base is added to create an overhang through the A-
tailing process. This A overhang permits adapters with only a 
single thymine base to pair DNA fragments. When performing 
RNA-seq, RNA must first be transcribed into cDNA. The 
fragmentation process can be carried out prior to or following 
cDNA synthesizing.
The addition of adapters occurs after fragmentation (or when 
it comes to library tagmentation preparation in conjunction 
with fragmentation). Adapters are permanently attached to 
the edges of DNA fragments. The adapters have multiple 
uses. They can transfer sequences into a flow cell, ensure 
compatibility with specific sequencing platforms, including 
barcodes (also known as indexes) to help identify the 
samples, and allow multiplexing in both target enrichment 
and sequencing. IDT has a range of options available in NGS 
adapters and indexing primers.
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You
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