# Ordering DNA

## IDT synthetic oligonucleotides

Much of our routine cloning work requires short single stranded DNA fragments known as oligonucleotides. More details coming soon!

## IDT gBlocks

We have [code](https://github.com/hsalis/Private-Code/blob/master/DNAc/shared/seq_assess.py) now that allows for checking of IDT gblock synthesis! Use it to aid in removal of unwanted motifs and features.

*Resuspending your gBlocks*

The dried down gBlocks Gene Fragment pellet can become displaced from the bottom of the tube during shipping.

1. Centrifuge the tube for 3−5 sec at a minimum of 3000 x g to pellet the material to the bottom of the tube.
2. Add ddH2O to the tube for your desired final concentration
3. Briefly vortex and centrifuge
4. (In table, TE buffer is supposed to be ddH2O)

## Resuspension volume of TE buffer (µL) for gBlocks Fragments synthesis scales

| Final Concentration (ng/µL) | 250 ng          | 500 ng | 1000 ng |
| --------------------------- | --------------- | ------ | ------- |
| 10                          | 25 µL           | 50 µL  | 100 µL  |
| 20                          | Not recommended | 25 µL  | 50 µL   |
| 50                          | Not recommended | 10 µL  | 20 µL   |

*Storing your gBlocks Gene Fragments*

gBlocks Gene Fragments can be stored in TE at −20°C for up to 24 months. If gBlocks Gene Fragments will be stored for less than 1 month, they can be resuspended in nuclease-free water instead of TE.

## Addgene Plasmids

Some plasmids are available at [Addgene](https://www.addgene.org/). Once you find the plasmid, message Dr. Salis to place the order. He has a PI account to place orders. See shipping material about how to obtain the plasmid DNA from the sample sent.

## CGSC Plasmids

Some plasmids are only available through the [E. coli Genetic Resources at Yale CGSC, The Coli Genetic Stock Center](https://cgsc2.biology.yale.edu/index.php). Go onto their website, find the plasmid, and the unique CGSC# identifiers. Send an email to [cgsc@yale.edu](https://github.com/reisalex/salis-lab-protocol-book/tree/453898c9360786eef221e6fffd8409c03a547e50/cgsc@yale.edu) requesting the plasmids. Make sure to include our lab address. Confirm with Dr. Salis in advance, and CC him. My emails in the past looked something like:

```
I'm with the Howard Salis lab at Penn State University. We'd like to request the following strains:
CGSC#: 12119 (Strain BW27783)
CGSC#: 7881 (Strain BW27784)
CGSC#: 7882 (Strain BW27786)

Please let me know what we need to do to continue forward with this request.

Our lab address is:
Howard Salis
Room 113 Tyson Building
Shortlidge Rd
University Park PA 16802
```

Make sure to follow the instructions on the shipping material about how to obtain the plasmid DNA!

## Oligo Pool Synthesis

Details coming soon!


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