Objective: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be use...Objective: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B. Methods: The total RNA from liver tissue with chronic hepa- titis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5′ end of the RNA transcript (SMART) technique and CDS III/3′ primer were used for first-strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to λTriplEx2 vector. Then λ phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. Results: The titers of unamplifed and amplified libraries were 1.94×106 pfu/ml and 1.49×109 pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1?2 kb in 64.29%, and 0.5?1.0 kb in 35.71%. Conclusion: A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed.展开更多
The character of Shylock in the Merchant of Venice is both intriguing and compelling. Hateful and greedy as he is on the onehand, yet suffered and pathetic on the other. Shakespeare is quite considerate in creating th...The character of Shylock in the Merchant of Venice is both intriguing and compelling. Hateful and greedy as he is on the onehand, yet suffered and pathetic on the other. Shakespeare is quite considerate in creating the character. This paper explores the possibili-ties behind Shakespeare, contradictory construction of Shylock.展开更多
A characterization of linear symplectic Runge-Kutta methods, which is based on the W-transformation of Hairer and Wanner, is presented. Using this characterization three classes of high order linear symplectic Runge-K...A characterization of linear symplectic Runge-Kutta methods, which is based on the W-transformation of Hairer and Wanner, is presented. Using this characterization three classes of high order linear symplectic Runge-Kutta methods are constructed. They include and extend known classes of high order linear symplectic Runge-Kutta methods.展开更多
基金Project (No. 30371270) supported by the National Natural Science Foundation of China
文摘Objective: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B. Methods: The total RNA from liver tissue with chronic hepa- titis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5′ end of the RNA transcript (SMART) technique and CDS III/3′ primer were used for first-strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to λTriplEx2 vector. Then λ phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. Results: The titers of unamplifed and amplified libraries were 1.94×106 pfu/ml and 1.49×109 pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1?2 kb in 64.29%, and 0.5?1.0 kb in 35.71%. Conclusion: A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed.
文摘The character of Shylock in the Merchant of Venice is both intriguing and compelling. Hateful and greedy as he is on the onehand, yet suffered and pathetic on the other. Shakespeare is quite considerate in creating the character. This paper explores the possibili-ties behind Shakespeare, contradictory construction of Shylock.
基金This work has been supported by the Swiss National Science Foundation.
文摘A characterization of linear symplectic Runge-Kutta methods, which is based on the W-transformation of Hairer and Wanner, is presented. Using this characterization three classes of high order linear symplectic Runge-Kutta methods are constructed. They include and extend known classes of high order linear symplectic Runge-Kutta methods.
