3258 pCMV5-anti-mIgGFc-rFc-His

Cloning of 3258 pCMV5-anti-mIgGFc-rFc-His

Purpose: Cloning of 3258 pCMV5-anti-mIgGFc-rFc-His

Date: 08, 15, 2018

Notebook: 

Page:

Clone ID: 3258

Experimenters: yijia

Results

Cloning

gene synthesis of 3258

template:

gene  synthesis

3258-1TGTCACGAATTCACAGGTGCAGCTGGTTGAATCTGGCGGAG

3258-2CGGCACAATACAGTCTCAGGCTTCCGCCAGGCTGAACCAGTCCTCCGCCAGATTCAACCA

3258-3CCTGAGACTGTATTGTGCCGCCAGCGGCAGAACCGACACAACATATGCCCTCGGCTGGTT

3258-4GATGCTGGCCACGAACTGTCTTTCTTTGCCAGGAGGCTGTCTGAACCAGCCGAGGGCATA

3258-5CAGTTCGTGGCCAGCATCACCTGGATCGGCGGAGCCACAAATTACGCCGCCTCTGTGAAG

3258-6TGAATGTGTTGGTGCCCAGGTCCTTGCTGATGGTGAATCTGCCCTTCACAGAGGCGGCGT

3258-7CTGGGCACCAACACATTCAACCTGCAGATGAACAGCCTGACACCTGACGACACCGCCGTG

3258-8TGAGCATTGGGCCACTGTCTAACGACGGCGGCAGCACAATAGTACACGGCGGTGTCGTCA

3258-9ACAGTGGCCCAATGCTCATCAGGGCGCCTACGATTATTGGGGCCAGGGCACACAAGTGAC

3258-10CTTGCTGGTACCGCTAGCACTAGACACGGTCACTTGTGTGCCCTGG

digest with EcoR1 + Nhe1 or EcoR1+ Kpn1

Vector digestion

Fastdigest  1 hour EcoR1+Kpn1

Vector is 3257

minprep of 3258

 Colony screen

3258 PCR Screening

Pick up#1,#3,#4,#6 to grow up o/n to do miniprep digestion

3258 Miniprep digestion

Failed, might be empty vector

9/3/2018, digest miniprep DNA with EcoR1-Kpn1

3258 PCR SCreening

3258 Miniprep digestion

EcoR1+Kpn1

Result intepretation/Summary

Conclusion