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29 buffer

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ϕ29 Buffer is a solution used for the storage and handling of the ϕ29 DNA polymerase enzyme. It is designed to maintain the stability and activity of the enzyme during storage and use.

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Lab products found in correlation

29 dna polymerase, by New England Biolabs (2 mentions) Typhoon 9400 phosphoimager, by GE Healthcare (2 mentions) Bovine serum albumin (bsa), by New England Biolabs (2 mentions)

2 protocols using 29 buffer

1

C-circle Assay Protocol for Telomere Length

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The method employed for the assay probing for C-circles was slightly modified from that performed in Henson et al.19 (link). Briefly, genomic DNA was prepared as above. Digested DNA was cleaned up by phenol-chloroform extraction and precipitation. DNA was diluted and measured using a Nanodrop spectrophotometer. Generally, 100 and 50 ng of DNA were used for each sample (10 μl). DNA was combined with 10 μl 0.2 mg/ml BSA (NEB), 0.1% Tween, 1 mM each dATP, dGTP and dTTP, 1X ϕ29 Buffer (NEB) and 7.5 U ϕ29 DNA polymerase (NEB) and incubated at 30 °C for 12 h then 65 °C for 20 min. Reaction products were diluted to 100 μl with 2XSSC and dot-blotted onto a 2XSSC soaked nylon membrane. DNA was UV-cross-linked onto the membrane, and then hybridized at 50°C with end-labeled 32P-(CCCTAA)4 oligo probe. Blots were washed, exposed and scanned using a Typhoon 9400 PhosphoImager (Amersham, GE Healthcare) and analyzed using ImageQuant Software.
Rivera T., Haggblom C., Cosconati S, & Karlseder J. (2016). A balance between elongation and trimming regulates telomere stability in stem cells. Nature structural & molecular biology, 24(1), 30-39.
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2

C-circle Assay Protocol for Telomere Length

Check if the same lab product or an alternative is used in the 5 most similar protocols
The method employed for the assay probing for C-circles was slightly modified from that performed in Henson et al.19 (link). Briefly, genomic DNA was prepared as above. Digested DNA was cleaned up by phenol-chloroform extraction and precipitation. DNA was diluted and measured using a Nanodrop spectrophotometer. Generally, 100 and 50 ng of DNA were used for each sample (10 μl). DNA was combined with 10 μl 0.2 mg/ml BSA (NEB), 0.1% Tween, 1 mM each dATP, dGTP and dTTP, 1X ϕ29 Buffer (NEB) and 7.5 U ϕ29 DNA polymerase (NEB) and incubated at 30 °C for 12 h then 65 °C for 20 min. Reaction products were diluted to 100 μl with 2XSSC and dot-blotted onto a 2XSSC soaked nylon membrane. DNA was UV-cross-linked onto the membrane, and then hybridized at 50°C with end-labeled 32P-(CCCTAA)4 oligo probe. Blots were washed, exposed and scanned using a Typhoon 9400 PhosphoImager (Amersham, GE Healthcare) and analyzed using ImageQuant Software.
Rivera T., Haggblom C., Cosconati S, & Karlseder J. (2016). A balance between elongation and trimming regulates telomere stability in stem cells. Nature structural & molecular biology, 24(1), 30-39.
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