pDNA Extraction and Quantification Class!

Description:
Ever wanted to make bacteria glow or create your own GMOs? This class will cover a few of the critical steps involved in many molecular biology projects. Learn firsthand how to isolate unique DNA from E.Coli containing special genes as well as measure how much DNA you produced. DNA like this could then be used in numerous downstream applications such as in agarose gels to isolate a gene or in transduction to other cells the genes. The class is meant as an introductory course into the field of genetics and molecular biology. It involves basic algebra and following directions to achieve desirable results. This class will consist of a five to fifteen minute introduction followed by hands on practice. If done correctly, each participant should end up with a micro centrifuge tube of plasmid DNA. While each process involved does not take long, inexperience and larger class sizes can greatly stretch things out.

When: Monday 17th at 1:00 PM - 6:00 PM
Where: Science
Cost: $15 to DMS

Safety Considerations:
!!!-NO COVID-19-!!!
IF YOU HAVE COVID-19, ANY SYMPTOMS OF COVID-19, OR COME INTO CLOSE CONTACT WITH THOSE WHO HAVE IT, DO NOT SIGN UP PLEASE!!!
IF YOU CAN SIGN UP, FACE MASKS MUST COMPLETELY COVER YOUR NOSE AND MOUTH!
SOCIAL DISTANCING IS REQUIRED DURING THE CLASS!
DO YOUR PART TO FLATTEN THE CURVE AND STOP THE SPREAD OF COVID-19!

Please wear closed toed shoes
Wear clothing that could get stained or thrown away (in the unlikely event anything does splash or spill)
No food or drink is allowed in the science area.
We will be dealing with E.Coli and other biological materials. You must strictly adhere to the sanitation policies outlined at the beginning of class.
For everyone’s safety, I will remove at my discretion any participant who does not follow any of the aforementioned rules. I cannot give refunds.

Link when it becomes available: https://calendar.dallasmakerspace.org/events/view/15896

Please let me know what you think in the thread below.

7 Likes

@NathanS Let me know if you need help with the class. I have experience working in the DIYBio molecular biology lab and tough similar classes in the past.

4 Likes

Looking forward to seeing the class! Looks really cool.

2 Likes

I can help if needed! I have degrees in biology and chemistry with research experience in stem cell and biochemistry.

Thank you for your support. The class shouldn’t be too technical as it will mainly be covering a miniprep and a Qubit BR dsDNA quantization kit. If you have any suggestions for improving miniprep yeilds or processes that would be great. I could only get about 11-18 ug/uL in about 100 ul the first few times I did it. Besides that, it is mostly just a process and learning how to use the tools. @dennis_o

Is it 100 ul of cells???
Are you using mini prep kit or are you following some DIY protocol?
What cells are you using?

I eluted the DNA into 100 ul of the 10 mM Tris-Cl, pH 8.5 buffer (EB buffer, QIAGEN). I mistyped earlier, I should have said “11-18 ug/mL” not “ul”. It is the QIAGEN spin miniprep kit here. I hope that clears it up a little.

I would say the concentration of plasmid is not bad. It is just not optimal.
How are growing the culture? If you grow culture too long then you can get too much dead cells debris. It could clog the miniprep. Ideally to use an overnight culture.
Also, what strain do you use? I’m not sure, it could be a strain with low plasmid copy number.

Registration is open now!