METHOD

Library preparation for metagenomic sequencing with Illumina

Krasnenko AYu1, Eliseev AYu2, Borisevich DI1, Tsukanov KYu1, Davydova AI1, Ilinsky VV1
About authors

1 Genotek Inc., Moscow

2 Moscow South-West High School No. 1543, Moscow, Russia

Correspondence should be addressed: Anna Krasnenko
Nastavnicheskiy per. 17, str. 1, pod. 14, Moscow, 105120; moc.liamg@oknensarkanna

About paper

Acknowledgements: the authors thank Daria Plakhina and Ivan Stetsenok of Genotek for their help and Sergey Glagolev of Moscow South-West High School No. 1543 for his valuable advice and comments.

Contribution of the authors to this work: Krasnenko AYu, Eliseev AYu — analysis of literature, research planning and implementation, data analysis and interpretation; Borisevich DI, Tsukanov KYu — bioinformatic analysis; Davydova AI — drafting of a manuscript; Ilinsky VV — research planning, scientific advisor. All authors participated in editing of the manuscript.

Received: 2017-04-12 Accepted: 2017-04-24 Published online: 2017-05-31
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Fig. 1. Agarose gel with PCR products after electrophoresis. The first well contains a DNA-size marker, the rest contain the samples. Primer dimers (PCR by-products) are marked with a black oval
Fig. 2. Sequencing quality control with Agilent Bioanalyzer 2100
Table 1. Universal primer pairs consist of a sequence complementary to the V3 and V4 regions of the 16S rRNA gene and a synthetic sequence complementary to Nextera and Truseq adapters
Table 2. Parameters of PCR for amplification of the V3 and V4 regions using universal primers for the Nextera adapter
Table 3. Nextera index primers
Table 4. Parameters of PCR for barcoding with Nextera primers
Таблица 5. Вид представления результатов биоинформатического анализа