Crm at 21:54, 19 April 2018

2018-04-19T21:54:32Z

New page

== Date:4/2/2018 ==

'''subclone of 984'''

Vector:1987

Insert:982

Size:957 bp

Enzymes:Xba1-Nhe1

[[File:IMG 0604.JPG|RTENOTITLE]]

'''redo subclone of 3152'''

Vector:3163

Insert: 3009pMiniT

enzymes:BsrGI+NheI

[[File:IMG 0606.JPG|RTENOTITLE]]

'''redo Subclone of 947'''

Vector:1727

Insert:947pMiniT

Size:1344 bp

[[File:IMG 0608.JPG|RTENOTITLE]]

== Date:4/3/2018 ==

'''redo PCR of 3127'''

new primer

3127-HindIII-R

3127-NheI-F2

size:403bp

extension time:30s

[[File:IMG 0612.JPG|RTENOTITLE]]

'''PCR of 986'''

F1:986-F1+986-R1

F2:986-F2+986-R2

Size:4kbp

Extension time:4min

[[File:IMG 0613.JPG|RTENOTITLE]]

== Date:4/4/2018 ==

'''miniprep digestion of 984_947_3152'''

#Name Enzymes Size:

1-4:984 XbaI+NheI                     957bp

5 :982(973-1pMiniT) XbaI+NheI

6-9:947 XbaI+NcoI-HF                1344bp

10:947pMiniT XbaI+NcoI-HF

11:947(from PCR cloning) XbaI+NcoI-HF

12-15:3152 BsrGI+NheI              371bp

16: 3162(Vector) BsrGI+NheI

[[File:IMG 0615.JPG|RTENOTITLE]]

'''3122 miniprep test digestion'''

1:3122-2

2:3122pJet1.2

enzymes:NdeI+SalI 265bp

3:3122-2

4:3122pJet1.2

Enzymes:EcoRV+SalI

[[File:IMG 0618.JPG|RTENOTITLE]]

'''938 miniprep test'''

1:938-1

2:938pJet1.2

Enzymes:EcoRV+XhoI

Size:552bp

3:938-1

4:938pJet1.2

Enzymes:NdeI+XhoI

Size:1071bp

[[File:IMG 0619.JPG|RTENOTITLE]]

'''939 miniprep test digestion'''

1:939-1 1-4 enzymes:NdeI+AgeI

2:939-2 1-4 Size: 421bp

3:939-4

4:939pJet1.2

5:939-1 5-8 enzymes:XhoI+AgeI

6:939-2 5-8 Size:355bp

7:939-4

8:939pJet1.2 Results:all of them have bands

[[File:IMG 0620.JPG|RTENOTITLE]]

== Date:4/5/2018 ==

'''subclone of 942'''

Size:716bp

Vector:947 pT7A3

Insert:942

[[File:Subclone942.JPG|RTENOTITLE]]

'''subclone of 986'''

Vector:1708

For 986 cloning

[[File:Vector1708forcloning986.JPG|RTENOTITLE]]

'''Subclone of 931'''

Vector:1987pT7A3

Insert:

F1:1174 bp

F2:986 bp

[[File:Subclone931.JPG|RTENOTITLE]]

'''824subclone'''

Size:~969bp

vector:1987

Insert:832pMiniT

[[File:Subclone824.JPG|RTENOTITLE]]

'''824subclone'''

Insert size:1743bp

Vector:1987pT7A3

[[File:Subclone3122.JPG|RTENOTITLE]]

'''Subclone 984'''

Vector:1987

Insert size:957 bp

[[File:Subclone 984.JPG|RTENOTITLE]]

'''Vector digestion for 3127'''

[[File:Subclone3127vector1987.JPG|RTENOTITLE]]

'''PCR of 944 F1+F2'''

Template:367, cut out band then combine

[[File:Pcr944F1+F2.JPG|RTENOTITLE]]

'''new way of 986 PCR'''

[[File:PCR986newway.JPG|RTENOTITLE]]

'''redo subclone of 3152'''

[[File:Subclone3152.JPG|RTENOTITLE]]

== Date:4/6/2018 ==

'''939 suclone'''

'''Size:780bp'''

'''Insert:939pJet1.2'''

'''Vector:1727 pT7A30'''

'''Enzymes:Nde1+Xho1'''

[[File:939subclone.JPG|RTENOTITLE]]

'''938 subclone'''

'''Size:1145bp'''

'''Insert:938pJet1.2'''

'''Vector:1727 pT7A30'''

'''Enzymes:Nde1+Xho1'''

[[File:938subclone.JPG|RTENOTITLE]]

'''3122 subclone'''

'''Size:1743bp'''

'''Insert:3122pJet1.2'''

'''Vector:1727 pT7A30'''

'''Enzymes:Nde1+Xho1'''

[[File:3122subclone.JPG|RTENOTITLE]]

'''3127 subclone'''

'''Size:408 bp'''

'''Insert:939pJet1.2'''

'''Vector:1727 pT7A30'''

'''Enzymes:Nde1+HindIII'''

[[File:3127subclone.JPG|RTENOTITLE]]

'''986 PCR'''

[[File:986pcr.JPG|RTENOTITLE]]

'''986 digestion as vector'''

'''Enzymes:HindIII+Xho1'''

[[File:986vectordigestion.JPG|RTENOTITLE]]

'''945 vector diegstion'''

'''Enzymes:Xba1+Nco1'''

[[File:945vectordigestion.JPG|RTENOTITLE]]

'''3166 PCR'''

'''Primer:3166-F+3166-R'''

'''Template:3017 pCEH0'''

'''Extension time:1min 30s'''

'''Size:1234bp'''

[[File:Pcr3166.JPG|RTENOTITLE]]

'''3152 subclone'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
|  Lane#
| Name
| Note
|-
| 1
| Undigested vector
| - control
|-
| 2
| digested vector
| - control
|-
| 3
| digested vector---3016 pCEH0
| for subclone
|-
| 4
| digested insert---3009pMiniT2
| for subclone
|}

[[File:3152subclone.JPG|RTENOTITLE]]

'''3129 digestion as vector'''

'''Enzymes:Sal1+BsrG1'''

[[File:3129vector diegstoinfor31653166.JPG|RTENOTITLE]]

'''945 subclone'''

'''Size:1326 bp'''

'''Enzymes:Xba1+Nco1'''

[[File:945subclone.JPG|RTENOTITLE]]

'''3165 PCR'''

[[File:PCRof3165.JPG|RTENOTITLE]]

'''944 PCR'''

'''Size:2334bp. might be wrong, because two bands in the gel.'''

[[File:PCRof944.JPG|RTENOTITLE]]

'''931 subclone'''

'''Vector:1518 pET28b digested with Nde1+Xho1'''

'''F1:1174 bp, digested with Nde1+HindIII'''

'''F2:986 bp, digested with HindIII+ Xho1'''

[[File:931subclone.JPG|RTENOTITLE]]

'''985 subclone'''

'''Vector:1987 pT7A3 digested with'''''' Nde1-Xho1'' '''Insert:983(973-2 pMiniT2) digested with Nde1+Sal1'''

'''Size:500 bp'''

[[File:985subclone.JPG|RTENOTITLE]]
[[Category:My ELN]]

Yijia LI 4 2 2018 - Revision history

Crm at 21:54, 19 April 2018