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From 2012.igem.org

I. Measure the Concentration of Miniprepped Plasmids

• Bring samples and Buffer EB bottle upstairs to Take3 plate reader
• Add 2 uL of each sample according to the following chart:

Blank (EB)	Blank
Blank	Blank
pBAV1K-T5-gfp I (15Jul12)	pBAV1K-T5-gfp II (15Jul12)
pBR322 I (18Jul12)	pBR322 II (18Jul12)
pBR322 III (18Jul12)	pBR322 IV (18Jul12)

• Use the program to blank the machine and measure sample concentrations

RESULTS

Concentration:

Plasmid	Concentration (ng/uL)	Approx. volume (uL)	Approx. total (ug)
pBAV1K-T5-gfp I	280.432	~46	12.90
pBAV1K-T5-gfp II	412.41	~42	17.32
pBR322 I	1.748	~500	0.874
pBR322-T5-gfp II	2.744	~500	1.372
pBR322-T5-gfp III	2.738	~500	1.369
pBR322-T5-gfp IV	1.749	~500	0.875

II. Ordered Updated PCR Primers from Keck

• Primers were ordered with the name/sequence in the following table:

Name	Tm (Tm overhang/Tm anneal)	Length (bp)	Sequence (overhangANNEAL)
cT5_pBf	74.16 (68.90/56.06)	66 (46/20)	5'-tgactctagatttgctttgtgagcggataacaattataatagattcAATTGTGAGCGGATAACAAT-3'
cT5_pBr	79.55 (75.05/56.42)	60 (44/16)	5'-gtaactgcaggagagcgttcaccgacaaacaacagataaaacgaAAGGCCCAGTCTTTCG-3'
cT5_p2f_2	74.20 (69.30/55.65)	66 (46/20)	5'-aattgtgagcggataacaattactagagaaagaggagaaatactagATGACCATGATTACGGATTC-3'
cT5_p2r_2	72.60 (63.89/55.41)	50 (30/20)	5'-tctccatacctcatcactcctcaatattacTTATTTTTGACACCAGACCA-3'
cT5_p3f_2	70.57 (62.42/56.57)	54 (32/22)	5'-caaaaataagtaatattgaggagtgatgaggtATGGAGAAAAAAATCACTGGAT-3'
cT5_p4r_2	81.11 (76.12/57.88)	59 (44/15)	5'-aaggcccagtctttcgactgagcctttcgttttatttgatgcctTCAGGTCGAGGTGGC-3'

• See Project Page for more info