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Heat shocked ‘blue’ ants.
Started trial ddRADseq library prep using 12 expendable Aphaenogaster samples that Nicole extracted last week - 3 ants from each colony.
Measured concentration using Qubit.
| Sample | Colony | ng/mL | Total DNA (ng) |
|---|---|---|---|
| 1 | EW205 | 424 | 1696 |
| 2 | EW110 | >600 | 2400 |
| 3 | 2012A1 | 291 | 1164 |
| 4 | DG23 | 580 | 2320 |
| 5 | 20130818-Mike | 349 | 1396 |
| 6 | EW20 QR | 352 | 1408 |
Total DNA = Qubit sample ng/mL * 100 * (1/1000 ul) * 40 ul
Combined Total DNA = 8 ug
For ‘real’ library would pool 24 - 48 samples, here only using 6 so used all DNA from each sample to ensure large volume of DNA in pooled sample for size selection and purification steps.
Double digest
50 ul rxn volume following NEB protocol
| Reagent | Volume (ul) |
|---|---|
| NlaIII | 1 |
| MluCl | 1 |
| DNA | 40 |
| CutSmart Buffer | 5 |
| H2O | 4 |
3 hr digestion at 37°C.
Included “C” Control sample of dH2O only.
Sent 2ul aliquot of each sample for Bioanalyer analysis of cutting efficiency.
Started PLOS Biology review
Cool video of Rob Dunn talking about making important scientific discoveries
This work is licensed under a Creative Commons Attribution 4.0 International License.