Crm at 01:15, 24 February 2018

2018-02-24T01:15:36Z

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Date:2/20/2018

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Template
| 2X PCR Mix
| Primers
|-
| 3027
| 2492-1,2,3
| Q 5 
| 3020-Xba1-Nhe1-F + 3027-2b-Not1-Age1-R 
|-
| 3076
| 3003 H-4,5,6
| Q 5
| 3076-myc-BsrG1-F + 3076-myc-Nhe1-Kpn1-R 
|}

[[File:PCR of 2492 3003 H.JPG|RTENOTITLE]]

Date:2/21/2018

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Template
| style="width: 156px;" | Restriction Enzymes
| Note
|-
| 1-5
| 3154 pMiniT2
| style="width: 156px;" | EcoR I
| wrong enzymes
|-
| 7,8,10
| 3021 pMiniT2
| style="width: 156px;" | EcoR I
| 
|-
| 11,14
| 3027 pMiniT2
| style="width: 156px;" | EcoR I
| 
|-
| Aim:
| Test if we succeed to put insert into pMiniT 2
| style="width: 156px;" | 
| 
|}

Digestion of 3154, 3021 or 3027 pMiniT2

[[File:Digestion of 3154 3021or 3027.JPG|RTENOTITLE]]

Date:2/22/2018

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| products
| Restriction Enzymes
| Note
|-
| 1,2
| 3010 L pMiniT2
| EcoR I
| 1
|-
| 3,4,5
| 3001 pMiniT2
| EcoR I
| 4
|-
| 6,7,8,9,10
| 3076 pMiniT2
| EcoR I
| 7
|-
| 11,12
| 3027 pMiniT2
| EcoR I
| 11
|-
| Destrisprtion
| We did miniprep and digested these products to test if we were sucessful.
| 1,4,7,11 to do sequencing
| 
|}

[[File:Digestion of 3010 L 3001 3027 3076.JPG|RTENOTITLE]]

Date:2/23/2018

Digestion Test for 3154 Restriction enzymes:BamH I+ Kpn I

[[File:DigestionTest for 3130.JPG|RTENOTITLE]]

Digestion for 3154 from miniprep

Restriction Enzyme:Kpn I+ BamH I

[[File:Digestion for 3154.JPG|RTENOTITLE]]

Result: good, ordered for sequencing;sequencing result is good too.

Digestion of Vector 3154 for 3129 ligation Cut by Kpn I + Not I

[[File:Digestion of Vector 3154 for 3129 .JPG|RTENOTITLE]]

We did ligation, but seems like the colonies are too much in a whole plate, so we planed to repeat but still picked up three colonies in this bad plate.

Digestion of Vetcor and Insert for 3129 ligation

[[File:Digestion of 3129 for ligation.JPG|RTENOTITLE]]

Digestion of Vector and Insert for 3027 Ligation

Restriction Enzymes: Nhe I+ Not I

[[File:Digestion of 3027 for ligation.JPG|RTENOTITLE]]

Because it looks like that the band of insert is not desired, so we repeated the digestion.

Another Digestion of Vector and Insert for 3027 Ligation

Restriction Enzymes:Nhe I+ Not I

[[File:Digestion of 3027 Ligation.JPG|RTENOTITLE]]

We got the similar band of the old one.So just cut the very light color band out,not sure, so cut both two light bands.

Digestion of insert from 3076 pMiniT2  for 3076 ligation

Restriction Enzymes:BsrG I+ Kpn I

[[File:Digestion of Insert 3076- pMiniT2 for 3076.JPG|RTENOTITLE]]

Not sure about the band, So planned to repeat next week.

PCR of 2493 to get the insert for 3021

2X PCR Mix:Platinum HiFi

Primers:3021-F2  + 3021-Age1-Xba1-R , Vector: BssHII - Nhe1

[[File:PCR of insert 2493 for 3021.JPG|RTENOTITLE]]

Good , so cut them and put into tube and add QG buffer
[[Category:My ELN]]

Yijia LI 2 19 2018 - Revision history

Crm at 01:15, 24 February 2018