1835-pT7A3-MBP-Taq-Muts-6His

Cloning of 1835-pT7A3-MBP-Taq-Muts-6His

Purpose: Cloning of 1835-pT7A3-MBP-Taq-Muts-6His

Date: 2013-9-17---2013=9-20

Notebook:0

Page:

Clone ID:1835

Experimenters: yingying

Results

cloning

PCR from 352 pET3a-Muts

Taq-Muts-F+Taq-Muts-HindIII-R

lane	1	2
	PCR product	1699 pT7A3-MBP-(E3C)
enzymes	Nde1 + HindIII

Colony Screening

Date: 2013-9-20

Make miniprep and digest with Nde1 + HindIII

<br>

Protein Purification

9/29/2013

Purification of Nde1 using 25 ml heparin HyperD column (XK16).

Dilute partial purified Nde1 with 50 mM Tris-HCl. Load to heparin column.

Elute with 250 ml linear gradient of 0.1 M to 0.6 M NaCl. Nde1 is eluted at 0.4 M NaCl.

Lane	Clone ID	Desc
1	1710	Nde1, Eluted from Heparin column
2	1710	Nde1, Eluted from Heparin column
3	1710	Nde1, Eluted from Heparin column
4	1813	FLP, 50 mM I wash from Ni column
5	1813	FLP, 300 mM I eluted from Ni, diluted
6	1710	Nde1, Eluted from Heparin column
7	1813	FLP, 300 mM I eluted from Ni, diluted
8	1813	FLP, heparin column FT
9	1813	FLP, 50 mM I wash from Ni column
10	1835	MBP-Taq-Muts-6XHis, partial purified from Ni column
11	1813	FLP heparin column elute
M		Fermentas PageRuler Plus
12	1813	FLP heparin column elute
13	1813	FLP heparin column elute