Thermo gel extraction and pcr cleanup kit

Manufactured by Thermo Fisher Scientific

The Thermo gel extraction and PCR cleanup kit is a laboratory product designed to efficiently purify DNA fragments from agarose gels and remove unwanted components from PCR reactions. The kit utilizes a silica-based membrane technology to selectively bind DNA, allowing for the removal of contaminants such as salts, primers, and unincorporated nucleotides.

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Lab products found in correlation

Dpni, by New England Biolabs (2 mentions)

Close spelling variants of this product

PCR Cleanup Kit Gel extraction kit Gel extraction

2 protocols using thermo gel extraction and pcr cleanup kit

Isolation of CMV and HC/LC-TBGH Fragments

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Final cassettes include CMV, and HC/LC-TBGH polyA fragments. To isolate these fragments, amplicons were first synthesized by PCR. PCR products were run on a 1% agarose gel and fragments of the correct length were extracted with Thermo gel extraction and PCR cleanup kit (ThermoFisher Scientific). Gel-extracted products were digested with DpnI (New England Biolabs) to further remove any possible contaminating plasmid. These fragment templates were then further amplified to create final stocks of cassette fragments.

Lima N.S., Musayev M., Johnston T.S., Wagner D.A., Henry A.R., Wang L., Yang E.S., Zhang Y., Birungi K., Black W.P., O’Dell S., Schmidt S.D., Moon D., Lorang C.G., Zhao B., Chen M., Boswell K.L., Roberts-Torres J., Davis R.L., Peyton L., Narpala S.R., O’Connell S., Wang J., Schrager A., Talana C.A., Leung K., Shi W., Khashab R., Biber A., Zilberman T., Rhein J., Vetter S., Ahmed A., Novik L., Widge A., Gordon I., Guech M., Teng I.T., Phung E., Ruckwardt T.J., Pegu A., Misasi J., Doria-Rose N.A., Gaudinski M., Koup R.A., Kwong P.D., McDermott A.B., Amit S., Schacker T.W., Levy I., Mascola J.R., Sullivan N.J., Schramm C.A, & Douek D.C. (2022). Primary exposure to SARS-CoV-2 variants elicits convergent epitope specificities, immunoglobulin V gene usage and public B cell clones. bioRxiv.

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Cassette Amplification and Purification

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Final linear cassettes include CMV, and HC/LC-TBGH polyA fragments. To isolate cassette fragments and introduce 15–20 base-pair overlaps, amplicons were first synthesized by PCR (CMV_FWD + CMV_TSO_REV, LC_FWD + TBGH_REV, HC_FWD + TBGH_polyA_REV). Crude PCR products were run on a 1% agarose gel and fragments of the correct size were extracted using the Thermo gel extraction and PCR cleanup kit (ThermoFisher Scientific). Gel-extracted products were digested with DpnI (New England Biolabs) to further remove any possible contaminating plasmid. These fragment templates were then further amplified with the same primers in large batches to create final working stocks of each cassette fragment. All primer sequences are listed in Supplementary Table 5.

Lima N.S., Musayev M., Johnston T.S., Wagner D.A., Henry A.R., Wang L., Yang E.S., Zhang Y., Birungi K., Black W.P., O’Dell S., Schmidt S.D., Moon D., Lorang C.G., Zhao B., Chen M., Boswell K.L., Roberts-Torres J., Davis R.L., Peyton L., Narpala S.R., O’Connell S., Serebryannyy L., Wang J., Schrager A., Talana C.A., Shimberg G., Leung K., Shi W., Khashab R., Biber A., Zilberman T., Rhein J., Vetter S., Ahmed A., Novik L., Widge A., Gordon I., Guech M., Teng I.T., Phung E., Ruckwardt T.J., Pegu A., Misasi J., Doria-Rose N.A., Gaudinski M., Koup R.A., Kwong P.D., McDermott A.B., Amit S., Schacker T.W., Levy I., Mascola J.R., Sullivan N.J., Schramm C.A, & Douek D.C. (2022). Primary exposure to SARS-CoV-2 variants elicits convergent epitope specificities, immunoglobulin V gene usage and public B cell clones. Nature Communications, 13, 7733.

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