Week 11 3/18-3/24: Trying transformation with a different plasmid

 Introduction

    After our transformation before spring break, we want to venture out into performing transformation with different plasmids in E.coli. One notable plasmid that we worked on this week was PW1 Cas9. At the same time, we are still retrying to pRad 1 but this time, during the extraction process, we will test with different amount of elusion buffer since we want more mass densities in our cells. The less elusion buffer we use, the more dense the cells are going to be.

pRad 1 Procedure and Results

    For extracting pRad 1, we plan to do three different amounts of elusion buffer: 10 ul, 20 ul, and 30 ul of elusion buffer. We will do 6 reactions, 3 being set #1 (10 ul, 20 ul, and 30 ul of elusion) and the other 3 being duplicates.

Here are the results taken from the nanodrop: 

               ug/ml    A260/A280    A260/A230

E1 (10ul)   17.5        2.00                1.21

E2  (20ul)  26.4        1.95                1.90

E3  (30ul)  44.8        1.86                1.77

E1.            17.1        2.29                1.33

E2.           13.5        2.06                0.57

E3.         7.1            2.19                1.04

Still, our values were still really low. After talking to Jonathon, he suggests we grew our E.coli a bit longer or pack more cells during step 1 in our plasmid extraction process. 


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