What is NGS Library Preparation How Does it Work

WHAT IS NGS LIBRARY PREPARATION? HOW DOES IT WORK? https://tekmatic.com

NGS (NEXT-GENERATION SEQUENCING) LIBRARY PREPARATION IS CONVERTING DNA SAMPLES INTO A FORM THAT CAN BE SEQUENCED ON AN NGS PLATFORM. THIS PROCESS INVOLVES SEVERAL STEPS, INCLUDING FRAGMENTATION, ADAPTER LIGATION, AMPLIFICATION, AND PURIFICATION. THE RESULTING LIBRARIES CAN BE USED FOR VARIOUS APPLICATIONS, INCLUDING GENOME SEQUENCING, GENE EXPRESSION ANALYSIS, AND DNA METHYLATION ANALYSIS. THIS BLOG WILL DISCUSS THE MULTIPLE STEPS INVOLVED IN NGS LIBRARY PREP AND THE IMPORTANCE OF THIS PROCESS IN MODERN GENOMICS RESEARCH.

HOW DOES NGS LIBRARY PREPARATION WORK? The first step in NGS library Prep is fragmentation, which involves breaking the DNA sample into smaller pieces. Several methods for fragmenting DNA include mechanical shearing, sonication, and enzymatic digestion. 03

The choice of fragmentation method depends on the specific NGS platform and the size of the DNA fragments desired. For example, shorter DNA fragments are often preferred for RNA-seq, while longer pieces may be more suitable for whole genome sequencing. After fragmentation, the DNA fragments are usually ligated to adapters. Adapters are short, synthetic oligonucleotides designed to bind specifically to the ends of the DNA fragments. These adapters serve several purposes.

FIRST, THEY ALLOW THE DNA FRAGMENTS TO BE CAPTURED AND AMPLIFIED DURING THE NEXT STEP OF LIBRARY PREPARATION. SECOND, THEY PROVIDE THE NECESSARY SEQUENCES FOR THE NGS PLATFORM TO RECOGNIZE AND BIND THE DNA FRAGMENTS DURING SEQUENCING. FINALLY, THEY MAY ALSO CONTAIN ADDITIONAL INFORMATION, SUCH AS BARCODES, WHICH CAN IDENTIFY THE SAMPLES OR DISTINGUISH BETWEEN DIFFERENT LIBRARIES IN A MULTIPLEXED SEQUENCING EXPERIMENT. ONCE THE DNA FRAGMENTS HAVE BEEN LIGATED TO ADAPTERS, THE RESULTING LIBRARIES ARE USUALLY AMPLIFIED THROUGH PCR (POLYMERASE CHAIN REACTION). IT ALLOWS MANY COPIES OF THE LIBRARIES TO BE PRODUCED, WHICH IS NECESSARY FOR THE LOW-INPUT NATURE OF NGS PLATFORMS.

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