32. Title: More DNA extraction of dnrp1 and WT strains#

32.1. Date#

15102020

32.2. Objective#

To have more SATAY DNA from WT and dnrp1 strains to further sequence .

32.3. Method#

  • Genomic extraction according Benoit protocol. See HERE

  • Use 2x more pellet , and upscaled the whole protocol to have more DNA to play with.

  • Pellet weights:

    • WT:

    • dnrp1_1:

    • dnrp1_2:

  • The final volume of DNA was :

    • WT: ~ 550uL

    • dnrp1: ~750uL

    • dnrp1 : ~750ul

32.4. Results#

A bit of smear due to shear in the DNA{width=50%}

  • Using Biorad Image Lab software to quantify the bands from gel, taking the 600bp band as a reference.

  • The 600bp band has 60ng of DNA in the 1kB GeneRuler ladder.

{width=90%}

  • The concentrations achieved with this method were:

Strain

Mean (ng/uL)

std

Significance of the std

Final Volume

Total DNA

WT

27.911019

1.084485

0.038855084

550

15ug

dnrp1_1

40.583937

1.576893

0.038855102

750

30ug

dnrp1_2

34.738095

1.349751

0.038855067

750

26ug

32.5. Conclusion#

  • I have enough DNA to continue for the digestion and ligation in order to send those samples for sequencing.