ScholarLens Back to search
2003•Di-Si Junyi Daxue xuebaoRequires access

Construction and characterization of human myeloma cell cDNA library

Gang Chen, Wanggang Zhang, Jie Fu, Fuyang Li, Xinping Liu, Libo Yao

Open publisher page 0 citations

Abstract

AIM: To construct human myeloma cell cDNA expression library. METHODS: Total RNA were extracted from human myeloma cell line HMy2 and mRNA were purified with Oligotex mRNA kit. 1 st and 2 nd strands of cDNA were synthesized through reverse transcription. After blunting the cDNA termini, the cDNA fragments were connected with Eco RI adapters, and the end of Eco RI adapters was phosphorylated. The cDNA smaller than 400 bp were removed by Sephacryl S400 spin column,the remaining fragments were ligated with the dephophrylated arms of λ gt11. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli Y1090 for titration. RESULTS: The HMy2 cell line cDNA library consisting of 1.5 ×10 6 recombinant bacteriophages was constructed for the first time. The average exogenous insert of the recombinants was about 1.4 kb. CONCLUSION: The constructed cDNA library can be used to screen target clones.

About this research paper

What this paper is about

AIM: To construct human myeloma cell cDNA expression library. METHODS: Total RNA were extracted from human myeloma cell line HMy2 and mRNA were purified with Oligotex mRNA kit. 1 st and 2 nd strands of cDNA were synthesized through reverse transcription. After blunting the cDNA termini, the cDNA fragments were connected with Eco RI adapters, and the end of Eco RI adapters was phosphorylated. The cDNA smaller than 400 bp were removed by Sephacryl S400 spin column,the remaining fragments were ligated with the dephophrylated arms of λ gt11. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli Y1090 for titration. RESULTS: The HMy2 cell line cDNA library consisting of 1.5 ×10 6 recombinant bacteriophages was constructed for the first time. The average exogenous insert of the recombinants was about 1.4 kb. CONCLUSION: The constructed cDNA library can be used to screen target clones.

Why it matters

A significance statement is not available in the OpenAlex record.

Key contribution

A contribution statement is not available in the OpenAlex record.

Method / approach

Method details are not available in the OpenAlex metadata.

Main findings

Findings are not separately available in the OpenAlex metadata.

Limitations

Limitations are not available in the OpenAlex metadata.

Applications

Application details are not available in the OpenAlex metadata.

Available abstract

AIM: To construct human myeloma cell cDNA expression library. METHODS: Total RNA were extracted from human myeloma cell line HMy2 and mRNA were purified with Oligotex mRNA kit. 1 st and 2 nd strands of cDNA were synthesized through reverse transcription. After blunting the cDNA termini, the cDNA fragments were connected with Eco RI adapters, and the end of Eco RI adapters was phosphorylated. The cDNA smaller than 400 bp were removed by Sephacryl S400 spin column,the remaining fragments were ligated with the dephophrylated arms of λ gt11. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli Y1090 for titration. RESULTS: The HMy2 cell line cDNA library consisting of 1.5 ×10 6 recombinant bacteriophages was constructed for the first time. The average exogenous insert of the recombinants was about 1.4 kb. CONCLUSION: The constructed cDNA library can be used to screen target clones.

Key concepts: Complementary DNA, cDNA library, Molecular biology, Reverse transcriptase, Biology, Recombinant DNA, Messenger RNA, RNA

Related papers

Related paper 1Related paper 2Related paper 3
Back to paper searchBrowse research topicsOriginal source
Construction and characterization of human myeloma cell cDNA library — Research Paper | ScholarLens