0.5 M EDTA, pH 8.0 (disodium ethylenediamine tetraacetate):

186.1 g Na2EDTA

Dissolve in approx. 400 ml ddH₂O, adjust pH to 8.0 with 10 N NaOH, and adjust to 1 liter final volume with distilled water

100 mM EDTA:

20 ml 0.5 M EDTA

80 ml ddH₂O

100 ml

95% ethanol/0.12 M NaOAc (ethanol/acetate):

95 ml 100% ethanol

4 ml 3 M NaOAc pH 4.5

1 ml ddH₂O

100 ml

5 mg/ml ethidium bromide (EtBr):

500 mg EtBr (Sigma E-8751)

ddH₂O to 100 ml

FE (formamide/EDTA): 5:1 (v/v) formamide:50 mM EDTA

10 ul ddH₂O

10 ul 100 mM EDTA

100 ul deionized formamide

make fresh

10X Fill-in/Kinase buffer:

(500 mM Tris-HCl, pH 7.6, 100 mM MgCl2, 10 mM DTT, and 50 ug/ml BSA in double distilled water)

5 ml 1 M Tris-HCl, pH 7.6

1 ml 1 M MgCl2

100 ul 1 M DTT

500 ul 1 mg/ml BSA

3.4 ml ddH₂O

10 ml

Fill-in Deoxynucleotide Preparation:

To make 4 ml of the fill-in nucleotides at a concentration of 0.25 mM of each nucleotide, combine the following:

500 ul PCR dNTPs (2 mM)

3500 ul ddH₂O

Aliquot this into 0.5 ml eppendorf tubes with 10 ul in each tube.

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