Cloning Membranipora and Terebratalia piwi, nanos, vasa, hedgehog, smoothened, runx

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Running a PCR overnight to clone the following genes:

Mm: Piwi1, Vasa, Runx
Tt: Hedgehog, Smoothened, Piwia, Piwib, Vasa, Nanos.

First time cloning Terebratalia genes.


Membranipora bands were ok and were purified directly (and ligated), but Terebratalia were too faint:

For this reason I ran a re-PCR for Terebratalia genes.


For the re-PCR I diluted the PCR product of the first reaction 1:20 and used as a template for a regular PCR (without changing any parameters). The bands this time were stronger, except for Piwia. Since unwanted bands appeared I had to cut the gel, extract, and purify the DNA.

Ligation was also set for these genes.


Heat shock transformation for all genes.