Genetic Engineering & Biotechnology News

JUL 2018

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GENengnews.com | Genetic Engineering & Biotechnology News | JULY 2018 | 11 so "1950s"—she and her team developed an agarose overlay method to pick isolated colonies. "We reasoned that we should be able to keep cells moist and hopefully aerated with a thin layer of molecular biology–grade agarose containing physiological concentra- tions of salts and glucose. We circled well- separated colonies with a marker, removed the medium, added warm melted agarose/ DMEM, waited a minute for the agarose to solidify, and then rapidly punched out iso- lated colonies of about 50 to 100 cells with sterile truncated pipette tips. Indeed, an ex- perienced operator can pick 48 clones from a 10-cm dish in 15 to 20 minutes—a far more rapid pace than using cloning rings," explains Dr. Lindberg. "Remember to supplement the isolated col- onies with fetal bovine serum if your cells are finicky," she cautions. "For some cell types, 50 cells alone will often die without added se- rum." She notes that the major problem oth- ers had with the technique is "in not correctly gauging the temperature of the sterile agarose, which must fall within a narrow range—warm and pourable, but not "hot hot." The Verified In-situ Plate Seeding (VIPS™) single-cell cloning system from Solentim is a unique "all-in-one" system which combines gentle seeding of single cells with immediate image confirmation of the single cell in the des- tination well. The VIPS system can generate a resultant output 96-well plate with high seed- ing efficiency rapidly; and due to the low-pres- sure, gentle nature of the dispensing method, the majority of these wells will go on to form colonies resulting in high cloning efficiencies. VIPS has to be initially purchased by cus- tomers for suspension CHO cells, but due to the gentle nature of the method, it should be well suited to a variety of other cell types which are normally damaged by FACS. VIPS is also incredibly easy to use and maintain, as compared to a FACS system. VIPS dispenses single cells directly onto the bottom of a dry well in a microtiter plate, confirms immediately the presence of a sin- gle cell, fills the wells with media, and then images the whole well—thereby eliminating the need for a separate whole-well imager. "Through the repeated droplet dispens- See Single-Cell Cloning on page 12 Drug Discovery YOU ARE NOT A ROBOT... SO DON'T ACT LIKE ONE FREE YOURSELF FROM ROUTINE PIPETTING www.integra-biosciences.com Making hands-free serial dilutions, reagent additions and sample reformatting very affordable for every lab. Compatible with all Integra's electronic pipettes from 4 to 16 channels for consistent results and unbeatable ergonomics. VOYAGER - Adjustable Tip Spacing Pipettes VIAFLO - Electronic Pipettes ASSIST PLUS Automating Multichannel Pipettes T h e V I P S s i n g l e - c e l l c l o n i n g s ys t e m f r o m Solentim is a small-footprint system that can sit inside a standard biosafety cabinet. Cloned colonies of Bon cells, a human carcinoid tumor cell model. An agarose overlay method was used to pick isolated colonies. The method was developed in the laboratory of Iris Lindberg, Ph.D. With this method, clonal cell lines from HEK, CHO, AtT-20, RIN, euro2A, and several other common and neuroendocrine cell lines have been derived. The black circles are early- appearing colonies, and the blue circles are later- appearing colonies. Iris Lindberg, Ph.D., Professor of Neurobiology and Anatomy, University of Maryland Medical School

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