Crm at 20:17, 19 April 2018

2018-04-19T20:17:57Z

New page

== Date:4/9/2018 ==

1.Pick up clonies

2.collect cells

3.redo subclone

== Date:4/10/2018 ==

'''Miniprep digestion of 3166pMiniT2_944pMiniT2_3127pMiniT2'''

Insert size:

3166pMiniT2:1234bp SalI+BsrGI

944pMiniT2 :2334bp NdeI+XhoI

3127pMiniT2:402bp Nde1-HindIII

[[File:IMG 0657.JPG|RTENOTITLE]]

'''944 subclone'''

[[File:944subclone.JPG|RTENOTITLE]]

'''3166 subclone'''

[[File:3166subclone.JPG|RTENOTITLE]]

'''Miniprep digestion of 945_986_944_3127'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Lane
| Size
| Enzymes
| Note
|-
| 945
| 1-4
| 1326bp
| Xba1-Nco1<br/>
| subclone
|-
| 986
| 5-8
| 774bp
| HindIII+XhoI
| ~4kbp
|-
| 944
| 9-12
| 2334bp
| NdeI+XhoI
| PCR
|-
| 3127
| 13-16
| 402bp
| Nde1-HindIII<br/>
| Gibson Mix
|}

[[File:IMG 0658.JPG|RTENOTITLE]]

'''Miniprep digestion of 984_942_824_3166'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Lane
| Size
| Enzymes
| Note
|-
| 984
| 1-4
| <br/>
| Xba1+NdeI
| Subclone
|-
| 942
| 5-8
| 716bp
| <div><div>Nco1-Xho1</div></div>
| <br/>
|-
| 824
| 9-12
| 969<br/>
| Nde1 + Xho1<br/>
| <br/>
|-
| 3166
| 13-16
| 1234bp
| Sal1+BsrG1<br/>
| PCR
|}

<div> [[File:IMG 0659.JPG|RTENOTITLE]]</div>
'''test for selecting correct subclone of 986_944_945_942_824'''

{| border="1" cellspacing="1" cellpadding="1" style="width: 500px;"
|-
| #
| Lane
| control
| Size
| style="width: 65px;" | Enzymes
| style="width: 195px;" | Note
|-
| 986
| 1-2
| Lane 3 1708 vector
| <br/>
| style="width: 65px;" | HindIII+XhoI<br/>
| style="width: 195px;" | undigested vector
|-
| 944
| 4-6
| Lane 7  944pMIniT2
| 2334bp
| style="width: 65px;" | NdeI+XhoI<br/>
| style="width: 195px;" | from PCR, '''No.4 might be right one'''
|-
| 945
| 8
| Lane 9  945pMiniT2
| 1326bp
| style="width: 65px;" | Xba1-Nco1<br/>
| style="width: 195px;" | '''good'''
|-
| 942
| 10-12
| Lane 13 942pJet1.2
| 716bp
| style="width: 65px;" | Nco1-Xho1<br/>
| style="width: 195px;" | '''good'''
|-
| 824
| 14-15
| Lane 16 832pMiniT2
| 969bp
| style="width: 65px;" | Nde1 + Xho1<br/>
| style="width: 195px;" | <br/>
|}

[[File:Redo test for miniprep.JPG|RTENOTITLE]]

== Date:4/11/2018 ==

'''Miniprep digestion of 3152_985'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Lane
| Size
| Enzymes
| Note
|-
| 3152
| 1-4
| 371bp
| BsrG1-Nhe1<br/>
| size is the same as digested vector,need to do sequencing
|-
| 985
| 5-8,  Lane 9 is +control
| 402bp
| Nde1+Kpn1
| good
|}

[[File:IMG 0667.JPG|RTENOTITLE]]

== Date:4/12/12018 ==

'''Miniprep digestion of 938_939_984'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Lane
| style="width: 34px;" | Enzymes
| style="width: 43px;" | Size
| Note
|-
| 938
| 1-4;5 is + control(sequenced 938pjet1.2)
| style="width: 34px;" | Nde1+Xho1
| style="width: 43px;" | 1145bp
| '''good'''
|-
| 939
| 6-9 ; 10 is + control(sequenced 939pJet1.2)
| style="width: 34px;" | Nde1+Xho1
| style="width: 43px;" | 780bp
| '''good'''
|-
| 984
| 11-14; 15 is + control(sequenced 982)
| style="width: 34px;" | Xba1+Nhe1
| style="width: 43px;" | 957bp
| need to redo
|}

[[File:Miniprepdigestion 938 939 984.JPG|RTENOTITLE]]

== Date:4/13/2018 ==

'''Miniprep of 944_3166_3127'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Lane
| Size
| Enzymes
| Result
| Note
|-
| 944
| 1-4; 5 is 944pminiT2
| 2334bp
| Nde1+Xho1
| Size is a little bigger than 2334 bp, need to do sequencing
| 944pMiniT2 is from PCR
|-
| 3166
| 5-9; 10 is 3166pminiT2
| 1234bp
| BsrG1+Sal1
| to do sequence
| 3166pMiniT2 is from PCR<br/>
|-
| 3127
| 11-14; 14 is 3127pMiniT2
| 402bp
| Nde1+HindIII
| to do sequence
| 3127pMiniT2 is from PCR
|}

[[File:IMG 0684.JPG|RTENOTITLE]]

== Date:4/14/2018 ==

'''Miniprep of 989_990_991_992'''

[[File:IMG 0685.JPG|RTENOTITLE]]

'''Miniprep of 993_995'''

[[File:IMG 0686.JPG|RTENOTITLE]]

'''Miniprep of 986'''
<div><div>#Ends Coordinates Length (bp) </div><div>1 HindIII-HindIII 1124-3468 2345 </div><div>2 HindIII-HindIII 93-1123 1031 </div><div>3 HindIII-HindIII 3469-92 893 </div></div>
[[File:IMG 0687.JPG|RTENOTITLE]]
[[Category:My ELN]]

Yijia LI 4 9 2018 - Revision history

Crm at 20:17, 19 April 2018