Method and kit for the generation of DNA libraries for massively parallel sequencing

A method for low-pass whole genome sequencing comprising the steps of:

• generating a plurality of barcoded, massively parallel sequencing libraries comprising the steps of:
• (a) providing a primary whole genome amplification (WGA) DNA library (pWGAlib) including fragments comprising a known 5′ sequence section (5SS), a middle sequence section (MSS), and a known 3′ sequence section (3SS) reverse complementary to the known 5′ sequence section, the known 5′ sequence section (5SS) comprising a WGA library universal sequence adapter, and the middle sequence section (MSS) comprising at least an insert section (IS), corresponding to a DNA sequence of the original unamplified DNA prior to WGA:
• (b) performing a PCR cycle consisting of a single round of PCR on the primary WGA DNA library using at least one first primer (1PR) comprising at least a first primer 5′ section (1PR5S) and a first primer 3′ section (1PR3S), the first primer 5′ section (1PR5S) comprising at least one first sequencing adapter (1PR5SA), and the first primer 3′ section (1PR3S) hybridizing to the known 3′ sequence section (3SS), so as to obtain a first primer extended WGA DNA library:
• (c) performing a PCR cycle consisting of a single round of PCR on the first primer extended WGA DNA library using at least one second primer (2PR) comprising a second primer 5′ section (2PR5S) and a second primer 3′ section (2PR3S), the second primer 5′ section (2PR5S) comprising at least one second sequencing adapter (2PR5SA) different from the at least one first sequencing adapter (1PR5SA), and the second primer 3′ section (2PR3S) hybridizing to the known 3′ sequence section (3SS), so as to obtain a first and second primer extended WGA DNA library:
• (d) amplifying by PCR the first primer extended by WGA DNA library and the second primer extended WGA DNA library using at least one third primer (3PR) comprising the first sequencing adapter (1PR5SA) and at least one fourth primer (4PR) comprising the second sequencing adapter (2PR5SA), so as to obtain an amplified first and second primer extended WGA DNA libraries, wherein the first primer 5′ section (1PR5S) further comprises at least one first sequencing barcode (1PR5BC), in 3′ position of the at least one first sequencing adapter (1PR5SA) and in 5′ position of the first primer 3′ section (1PR3S) and/or the second primer 5′ section (2PR5S) further comprises at least one second sequencing barcode (2PR5BC), in 3′ position of the at least one second sequencing adapter (2PR5SA) and in 5′ position of the second primer 3′ section (2PR3S);
• providing the plurality of barcoded, massively-parallel sequencing libraries and pooling samples obtained using different sequencing barcodes (BC);
• and
• sequencing the pooled library.