Does anyone have experience cloning 10kb fragment?

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Venkata Giridhar Poosarla

Follow the procedure from Gruber; I got success in cloning 10kb fragment.

Ayat Ibrahiem Esmaeel Al-Laaeiby

Thanks a lot

Ji Shi

if you know the sequence of the 10kb fragment, you can choose the restricted sites in the middle position, for example 4kb and 6kb ,then use overlap PCR ,The overlabed district should be 100bp or more ,after PCR, use ligase link the two fragments and clone the 10k to vector.