Team:UCLA/Notebook/Spider Silk Genetics/17 April 2015

iGEM UCLA





4/17/2015

4/17/2015


1 PCR amplification of pSB1C3

We set up a 50 uL reaction using the following protocol from Julian’s notebook.

Volume (uL)
5x Q5 Buffer 10
10 mM dNTPs 1
10 uM For (SB-prep 2EA) 2.5
10 uM Rev (SB-prep 3P1) 2.5
Template (Spycatcher from Julian)1
Q5 Polymerase .0.5
ddH2O 32.5
Total 50

98 C 30 sec
98 C 10 sec
66 C 20 sec
72 C 30 sec
repeat from step 230 x
72 C 2 min
12 C hold

2 Results

∙ Cast a 1% gel to visualize.

PIC

Figure 1: PCR amplification of pSB1C3

∙ Gel extraction (Qiagen) yielded 36.47 ng/uL, A 260/280: 1.91.