Nano-Drop

 Introduction

    This week was preparation leading into next week's project, ethanol precipitation. Ethanol precipitation is used for concentrating and de-salting nucleic acid (DNA or RNA). This week we were running our samples through gels and then extracting our samples once they've run through the gels. We started with a different variation of our usual 1% gel, this time we used a 1.5% gel. For this, we needed 30 mL of 1 time TAE and 450 mg of agarose powder. With the addition of an extra 5 seconds in the microwave and 150 mg of agarose powder, our gel is made. While we were running our gels, 

    We set the Nano-Drop to Double Stran DNA and tested the samples. 

The reason there are 2 pictures is that I ended the experiment by accident....

    By the time we finished on the Nano-Drop our gels had finished and it was time to place them over the UV light and extract the sample. 

    We measured each 2mL tube that the samples would be placed in for future calculations. We took the empty weight, then the weight with the sample. Once we had the weight of our gel, we could start with the extraction process. In between each step, we centrifuged the samples for one minute, then discarded the supernate.

Weight of the samples extracted

Right Flank 1	94.1 mg
Right Flank 2	95.2 mg
Right Flank 3	150.7 mg
Right Flank 4	148.2 mg

Amount of Buffer QG added (3 times volume of sample)

Right Flank 1	285 micro-liters
Right Flank 2	285 micro-liters
Right Flank 3	450 micro-liters
Right Flank 4	445 micro-liters

Amount of Isopropanol added (1 times volume of sample)

Right Flank 1	94 micro-liters
Right Flank 2	95 micro-liters
Right Flank 3	150 micro-liters
Right Flank 4	148 micro-liters

     After a few rounds of centrifuging, we had our finished product. We then took these samples back to the nano-drop to compare the results. 

Double-Stranded DNA

	Ng/Micro-liter	A260/A280	A260/A230
Right Flank 1	25.9	1.97	0.07
Right Flank 2	22.7	1.91	0.05
Right Flank 3	25.2	2.04	0.04
Right Flank 4	23.3	2.01	0.06

UV Visibility

	280	260	230	225
Right Flank 1	.27	.52	7.94	12.52
Right Flank 2	.24	.46	8.99	14.16
Right Flank 3	.26	.50	16.32	22.91
Right Flank 4	.25	.47	8.88	13.67

    From the results, ideally, we would like to have our A260/A280 as close to 1.80 for our DNA. As shown from the results, we are still off by a decent amount. But next week we continue our efforts in transforming Deinococcus Radiodurans.