3230 pEPD-DP47-DPL16

Cloning of

Purpose:  

Date: 2018-6-22

Notebook: 

Page:

Clone ID: 3230

Experimenters: Paul 

Results

PCR

Date: 6/21/2018

By: Paul

template:

primers:

Gel:

Cloning

By: Paul

Vector:  

Ligation: 30 min with T4 DNA ligase

Competent cells: NEBTurbo

Colony PCR Screening 

~ 

Restriction Digestion

Date: 6/~/2018

By: Paul

http://www.excellgen.net/eln/images/8/8c/3208_3231_3230_digestion.JPG

3230 and 3231 in pMiniT2, digest with EcoR1

Sequencing

'

Fix 3030

Insert: 3230 in pEPD, vector is 3208 pEPD-FW, yield 3230 Fix in pEPD

ligate and transform

Restriction Digestion

BamH1 + Xma1

both clone # 2 and 3 has 1 C missing, so the clone from 3230 in pEPD is not good

6/25/18

6/26/2018

3230 fix #4 is parental (3208)--result also confirmed by sequencing, it could not be cut by ApaI (no GGGCCC site) . 

sequencing: both 3230 fix #2 and 3230 fix #3  has 1 C missing

Redo fixing

Result intepretation/Summary

sequencing result: new 3230 clone #9 is good