1023-pET8XHis-AvPAL

Cloning of 1023-pET8XHis-AvPAL

Purpose: Cloning of 1023-pET8XHis-AvPAL

Date: 2012-3-22--2012-3-28

Notebook:0

Page:

Clone ID:1023

Experimenters: yingying

Results

cloning

Date:2012-3-22

AvPAL-1-1  to AvPAL-1-16

AvPAL-2-1  to AvPAL-2-18

AvPAL-3-1  to AvPAL-3-14

PCR 3 fragments and then using Gibson mix to join them

 

Digest:

	band 5	band 6
	PCR product	pET8XHis 286
enzymes	Nco1+ Xho1

Colony Screening

Date: 2012-3-26

PCR screen  with T7 F+AvPAL-3-14

no band

Result intepretation/Summary

• <br>

Conclusion

<br>

<br>

<br>

Cloning of 1023-pET8XHis-AvPAL

Purpose: Cloning of 1023-pET8XHis-AvPAL

Date: 2012-3-30--2012-4-2

Notebook:

Page:

Clone ID:1023

Experimenters: yingying

Results

cloning

Date:2012-3-22

fragment 1:AvPAL-1-1  to AvPAL-1-16

fragment 2:AvPAL-2-1  to AvPAL-2-18

fragment 3:AvPAL-3-1  to AvPAL-3-14

PCR 3 fragments and then fragment 1+fragment 2+fragment 3 as template

                                        AvPAL-1-1+AvPAL-3-14 as primers

digest :

	band	band
enzymes

Colony Screening

Date: 2012- 4-2

no insert

• 
  

Conclusion

<br>

Cloning of1023-pET8XHis-AvPAL

Purpose: Cloning of 1023-pET8XHis-AvPAL

Date: 2012-4-4--2012-4-6

Notebook:

Page:

Clone ID:1023

Experimenters: yingying

Results

cloning

Date:2012-4-4

fragment 1:AvPAL-1-1 to AvPAL-1-16

fragment 2:AvPAL-2-1 to AvPAL-2-18

fragment 3:AvPAL-3-1 to AvPAL-3-14

PCR 3 fragments and then fragment 1+fragment 2+fragment 3 as template

AvPAL-1-1+AvPAL-3-14 as primers

Digest:

	band 1	band 2
	PCR product	pET8XHis 286
enzymes	Nco1+ Xho1

Colony Screening

Date: 2012-4-6

Result intepretation/Summary

• &lt;br&gt;

Conclusion

&lt;br&gt;

&lt;br&gt;

<br>

Cloning of 1023-pET8XHis-AvPAL

Purpose: Cloning of 1023-pET8XHis-AvPAL

Date: 2012-4-4--2012-4-25

Notebook:

Page:

Clone ID:1023

Experimenters: yingying

Results

cloning

Date:2012-4-4

fragment 1:AvPAL-1-1  to AvPAL-1-16

fragment 2:AvPAL-2-1  to AvPAL-2-18

fragment 3:AvPAL-3-1  to AvPAL-3-14

PCR 3 fragments and then fragment 1+fragment 2+fragment 3 as template

                                        AvPAL-1-1+AvPAL-3-14 as primers

digest :

	band 3	band 4
	PCR product	pET8XHis-PKAalpha
enzymes	Nco1+ Xho1

Colony Screening

Date: 2012- 4-10

clone 3,5,6,7,8 may be right

subclone from 1023-3( insert ) to 1023-8 (vector), digest with Nco1+EcoR1

Date:2012/6/13

lane	1	2
plasmid	1236 pCDNA3.1-AvPAL-2	1023-1
enzymes	BamH1+EcoR1

6/15/2012

digest miniprep with EcoR1 + Bamh1

2012-6-20

digest detect :BamH1+EcoR1

2012-6-21

lane	1	2
	1236pCDNA3.1-AvPAL-2	1023-1
enzymes	BamH1+EcoR1

2012-6-25

digest detect:BamH1+EcoR1

2012-8-3

digest with:Nco1+Xho1

clone 1.4.6 right

• 
  

Conclusion

<br>