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From 2010.igem.org

Contents 1 146. labday 11.10.2010 1.1 cloning of lITR_CMV_betaglobin and lITR_phTERT_betaglobin into pSB1C3_CD 2 147. labday 12.10.2010: Example Example Example Example Example 3 148. labday 13.10.2010 4 149. labday 14.10.2010 5 150. labday 15.10.2010 6 151. labday 16.10.2010 7 152. labday 17.10.2010 8 153. labday 18.10.2010 9 154. labday 19.10.2010 10 155. labday 20.10.2010

146. labday 11.10.2010

cloning of lITR_CMV_betaglobin and lITR_phTERT_betaglobin into pSB1C3_CD

Investigator: Stefan

Comment: In order to test our GOI with another promotor, lITR_phTERT_beta-globin_mGMK_TK30 needs to be cloned upstream of hgH_rITR.

Vector name:
pSB1C3_CD clone 1 (P???)
pSB1C3_CD clone 2 (P???)

Insert name:
pSB1C3_lITR_CMV_beta-globin (P729)
pSB1C3_lITR_phTERT_beta-globin (P730)

Digestion:

components'	volume CD clone 1 + 2 /µl	volume P729 /µl'volume P730 /µl
DNA	6	14	6
BSA (10x)	2	2	2
Buffer 4 (10x)	2	2	2
Enzyme EcoI	1	1	1
Enzyme XbaI	1	-	-
Enzyme SpeI	-	1	1
H2O	8	-	8
Total volume (e.g. 15,20,25,30 µl)	20	20	20

Gel:
0,5 g Agarose, 50 ml TAE (1%), 3 µl GELRED , at 115 Volt

File:New.jpg

Gel extraction:
Was performed according to protocol.

T4 Ligation:

ligation name	728 + 670
volume of vector	2,35
volume of insert	5,65
T4 ligase buffer (10x)	1
T4 ligase	1

Transformation:
Was performed according to standard protocol using BL21 cells.

147. labday 12.10.2010: Example Example Example Example Example

148. labday 13.10.2010

149. labday 14.10.2010

150. labday 15.10.2010

151. labday 16.10.2010

152. labday 17.10.2010

153. labday 18.10.2010

154. labday 19.10.2010

155. labday 20.10.2010

=> Go to Labjournal October part 3 (labday 156 - 166 )