Laboratory Notebook: Group Meetings/Monoclonal; CSHL, 1988 - 1994

Display Title

Laboratory Notebook: Group Meetings/Monoclonal; CSHL, 1988 - 1994

Source

box: 07 folder: 04-05 (text)

Extent

2 Folders

Description

This is a notebook within a series on Greider's experiments and her contribution to group meeting. The notebook begins with a discussion of using phenol to extract DNA. Greider had dropped the phenol in a reducing agent and the placed the mixture into a tube. After creating the necessary reaction, she extracted the phenol, spun it and the re-extracted the contents. After extracting, Greider washed the resulting pellet. Then Grieder ran the results of the gel through a minigel of an enzyme combination of Bam and RI. Then began mixing together a pre-absorbed phage and E.coli- followed the growth, and graphed the result.

Grieder prepared another experiment by making buffer solutions for E. coli. She made three in total: KOAC Buffer, RNase Buffer, Chloroform Buffer. The notebook also included a diagram of certain plasmids, pUC and M13, with restriction enzyme. The specific restriction enzymes that Grieder used on the plasmids were: Acc1, P3t1, Hinc2, Tag.

After using enzymes on the plasmids Greider began to grow stock cultures of the phage and prepped the E.coli by titrating it. Also Grieder prepped two flasks of cells for the following days experiment. Greider inoculated the stock growth cell and placed it in shaker thereby infecting the cells. By infecting them Greider allowed the cells to incubate and harvest by spinning them together.

Greider made a mini prep, one with lysozymes and the other without lysozymes. Greider then purified the Bam/RI fragment of the plasmid from earlier and produced gels from both the Bam and RI digests. Greider then concentrated the DNA from the digest into a dried pellet and added an enzyme to it. Greider then poured the DNA into agarose gel and cut the DNA with a specific band using glass beads. However, the bands were too faint to take a picture. Greider conducted a series of transformation experiments, and placed frozen component cells in Glycerol. The process was quite simple as Grieder thawed and ligated the cell and then screened each colony on each plate.

Scope and Contents

This is a notebook within a series on Greider's experiments and her contribution to group meeting. The notebook begins with a discussion of using phenol to extract DNA. Greider had dropped the phenol in a reducing agent and the placed the mixture into a tube. After creating the necessary reaction, she extracted the phenol, spun it and the re-extracted the contents. After extracting, Greider washed the resulting pellet. Then Grieder ran the results of the gel through a minigel of an enzyme combination of Bam and RI. Then began mixing together a pre-absorbed phage and E.coli- followed the growth, and graphed the result.

Grieder prepared another experiment by making buffer solutions for E. coli. She made three in total: KOAC Buffer, RNase Buffer, Chloroform Buffer. The notebook also included a diagram of certain plasmids, pUC and M13, with restriction enzyme. The specific restriction enzymes that Grieder used on the plasmids were: Acc1, P3t1, Hinc2, Tag.

After using enzymes on the plasmids Greider began to grow stock cultures of the phage and prepped the E.coli by titrating it. Also Grieder prepped two flasks of cells for the following days experiment. Greider inoculated the stock growth cell and placed it in shaker thereby infecting the cells. By infecting them Greider allowed the cells to incubate and harvest by spinning them together.

Greider made a mini prep, one with lysozymes and the other without lysozymes. Greider then purified the Bam/RI fragment of the plasmid from earlier and produced gels from both the Bam and RI digests. Greider then concentrated the DNA from the digest into a dried pellet and added an enzyme to it. Greider then poured the DNA into agarose gel and cut the DNA with a specific band using glass beads. However, the bands were too faint to take a picture. Greider conducted a series of transformation experiments, and placed frozen component cells in Glycerol. The process was quite simple as Grieder thawed and ligated the cell and then screened each colony on each plate.

Preferred Citation

Laboratory Notebook: Group Meetings/Monoclonal; CSHL, 1988 - 1994. Carol Greider Collection, Cold Spring Harbor Laboratory Archives Digital Repository. 88-1525513. Update 2025-03-18.

Credit Line

Courtesy of Cold Spring Harbor Laboratory Archives.

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Level

file

Collection

Carol Greider Collection

Resource ID

CWG

Reference ID

0d4387de01f800bfefebdb9ccecae7e5

Finding Aid Link

Laboratory Notebook: Group Meetings/Monoclonal; CSHL