Excellgen at 20:15, 4 May 2015

2015-05-04T20:15:12Z

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== '''Cloning of 2423-pEC-pvuRts1I''' ==

'''Purpose:''' Cloning of 2423-pEC-pvuRts1I

Name: Xin

'''Date: '''2015 April

'''Results'''

Gene sysnthsis in two sections: 1-1 --- 1-10 ; 1-11- 1-20

The PCR about 450bp is what we interested.

[[File:2423, 2436 PCR 4-9-11.JPG|436x185px|RTENOTITLE]]

  

Gel purified the inteserted PCR product, and put togeter to do the big PCR.

[[File:2423 2436 PCR 4-9-15.JPG|382x223px|RTENOTITLE]]

The PCR product is about 900bp. Purifed the PCR product from gel. and digest by Nde1/xhoI

Also digest 2015 PEC2. ligation and transformation

Screening:

[[File:2423,2412 screening 4-17-15.JPG|242x180px|RTENOTITLE]]

Select colony#5 to sequence on 4/20/2015

== '''Fix:''' ==

Colony#5 sequencing result have one nucleotide was wrong.

First PCR fragments: Use oligos 2423-PvuRts1I-1, and -4: 160bp

Second PCR fragements: use oligos: 2423-PvuRts1I-5, and -20: 620bp

[[File:2423,2436,2421 PCR.JPG|364x152px|RTENOTITLE]]

Join two fragments together. 

[[File:2436, 2423, 2421 PCR.JPG|451x168px|RTENOTITLE]]

== '''''Screening''''': ==

[[File:1916 2423 2421 screening.JPG|475x152px|RTENOTITLE]]

#2 and 3 are positive colonies.

== '''Conclusion''' ==

== '''Result intepretation/Summary''' ==

== '''Conclusion''' ==
[[Category:Cloning]]

2423-pEC-pvuRts1I - Revision history

Excellgen at 20:15, 4 May 2015