PacBio HiFi de novo Sequencing

DNA Extraction and Quality Control

CLR sequencing HMW gDNA extractions methods

1) Sigma GenElute™ Bacterial Genomic DNA Kits
For Bacterial Genomes great results have been achieved using

2) Phenol/Chloroform
Absolutely make sure Phenol is fresh and not oxidised (even closed it should not be older than 3 months)
Essential to perform DNA cleanup with EtOH precipitation
EtOH precipitation with NaAc ph5.2
At least 3 washes with 70% EtOH at biggest volume possible

3) Qiagen Gentra® Puregene® kit (100-200 kb)

4) Promega wizard genomic DNA
Similar to Phenol/Chloroform but faster. Good results on our Sequel have been achieved isolating HMW DNA from blood, using this kit

5) Qiagen Genomic Tips
Produces pure high molecular weight DNA
Requires large amount of starting material

6) Qiagen® MagAttract® HMW lit (100-200 kb)

7) Plant DNA isolation Protocol with Qiagen Genomic tips + CTAB

8) The "Mayjonade" protocol

Additional resources

Recommendations regarding DNA extractions for PacBio Sequencing

Created by patrig. Last Modification: 2021-07-14 11:26 by patrig.