Prepared by Akram Tariq Sial
Overview: This sample was prepared according to manufacturer's recommendations. [Add link to illumina protocol]
1µg of 98.0 ng/µl genomic DNA was sheared using either Covaris or a Nebulizer. Lanes 3&4: Banana Slug DNA sheared using Covaris. Lanes 5&6: Banana Slug DNA sheared using a Nebulizer. Both were cut between 250-375 basepairs.
Bioanalyzer results of shearing distributions and final amplified libraries. Covaris and Nebulizer shearing are in lanes 1 and 2 respectively. Amplified and size selected libraries for Nebulizer and Covaris in lanes 3 and 4 respectively.
The results indicate the final nebulizer library ranges from 202-320bp in distribution. The final covaris library ranges from 225-263bp in length.
Each stage indicates a different distribution in the length of the paired-end reads. The original size selection cut indicates a fragment size of 250-375. However, the bioanalyzer results indicate the final size selected fragment is much shorter at 200-320 bp for a final distribution between 100-230 bp.
Each lane contained a different concentration of starting material used in bridge amplification.
|Lane||Starting amount of library and library type|
|Lane 1||8 picoMoles Nebulized Library|
|Lane 2||10 picoMoles Nebulized Library|
|Lane 3||12 picoMoles Nebulized Library|
|Lane 4||6 picoMoles phix control*|
|Lane 5||8 picoMoles Covaris Library|
|Lane 6||10 picoMoles Covaris Library|
|Lane 7||12 picoMoles Covaris Library|
|Lane 8||14 picoMoles Covaris Library|
These different starting quantities of DNA are reflected in the number of clusters generated and the total reads outputted for each lane. Additionally, a different distribution of distance for the paired-end reads are different for lanes 1-3 and 5-8
*Data from lane 4 of the illumina run SHOULD NOT be used for assembly